Sickle cell disease treatment utilizing omega-3 fatty acids

ABSTRACT

Described herein are compositions including at least one omega-3 fatty acid (either in the triglyceride, ester or free fatty acid ester form) and at least one surface active agent; wherein the compositions form micelles when in contact with an aqueous medium. Also provided are methods of administering to a subject a composition including at least one omega-3 fatty acid (either in the triglyceride, ester or free fatty acid ester form) and at least one surface active agent, wherein the compositions form micelles when in contact with an aqueous medium, and the bioavailability of the omega-3 fatty acid is substantially independent of a food effect. The compositions are useful for treating certain disease states which may include (1) malabsorption syndromes, (2) primary sclerosing cholangitis (PSC), (3) non-alcoholic fatty liver disease (NAFLD), (4) sickle cell disease (SCD), (5) age-related macular degeneration (AMD), and (6) neurodegenerative disease, including, Parkinson&#39;s Disease (PD), Alzheimer&#39;s Disease (AD), Amyotrophic Lateral Sclerosis (ALS or Lou Gehrig&#39;s Disease), Epilepsy, Bi-polar Syndrome, traumatic brain injury, peripheral neuropathy, and Multiple Sclerosis (MS). Described are also various dosage forms for administering the compositions and use of the compositions in functional foods. Provided herein are also kits with instructions for their administration.

CROSS REFERENCE TO RELATED APPLICATIONS

This application is a continuation of U.S. patent application Ser. No.16/655,643 filed on Oct. 17, 2019.

U.S. patent application Ser. No. 16/655,643 is a continuation of U.S.patent application Ser. No. 15/605,972 filed May 26, 2017.

U.S. patent application Ser. No. 15/605,972 is a continuation of U.S.patent application Ser. No. 15/180,430 filed Jun. 13, 2016.

U.S. patent application Ser. No. 15/180,430 is a continuation ofInternational Patent Application No. PCT/US15/54933, filed Oct. 9, 2015,which claims priority from U.S. Provisional Patent Application Nos.62/062,638 filed Oct. 10, 2014; 62/062,643 filed Oct. 10, 2014;62/062,646 filed Oct. 10, 2014; 62/062,652 filed Oct. 10, 2014;62/062,634 filed Oct. 10, 2014; and 62/062,651 filed Oct. 10, 2014.

FIELD OF THE INVENTION

This invention relates to compositions including fatty acids, either inthe triglyceride, ester, or free fatty acid forms, in combination withat least one surface active agent, and to the use of such compositionsfor the treatment of patients afflicted with various disease states.More particularly, this invention relates to self-micellizingcompositions including at least one omega-3 fatty acid, which may beselected from the group consisting of hexadecatrienoic acid, α-linolenicacid, stearidonic acid, eicosatrienoic acid, eicosapentaenoic acid,heneicosapentaenoic acid, the omega-3 isomer of docosapentaenoic acid,also known as clupanodonic acid, docosahexaenoic acid,tetracosapentaenoic acid, and tetracosahexaenoic acid, in combinationwith a surface active agent composition effective to spontaneously formmicelles with said fatty acids upon contact with an aqueous media. Incertain embodiments, the self-micellizing compositions may furthercontain an omega-6 fatty acid (either in the triglyceride, ester, orfree fatty acid form), for example arachidonic acid (ARA), linoleic acid(LA), gamma-linolenic acid (GLA) or the omega-6 isomer of DPA, alsoknown as osbond acid. Most particularly, this invention relates to thetreatment of various disease states by administering to a patient inneed thereof the aforementioned compositions or pre-formed micellesproduced therefrom.

In certain embodiments, the disease states may include (1) malabsorptionsyndromes, such as short bowel syndromes, which encompass a number ofdifferent clinical entities that may result in chronic diarrhea,abdominal distention, and failure to thrive; (2) primary sclerosingcholangitis (PSC), which is a chronic cholestatic liver diseasecharacterized by progressive inflammatory and fibrotic destruction ofthe intrahepatic and/or extrahepatic bile ducts; (3) non-alcoholic fattyliver disease (NAFLD), which is characterized by increased hepatic fataccumulation in individuals not consuming excessive alcohol andrepresents a spectrum of disease ranging from ‘simple’ steatosis tonon-alcoholic steatohepatitis; (4) sickle cell disease, which is a groupof blood disorders that affects hemoglobin, the molecule that deliversoxygen throughout the body via red blood cells; (5) age-related maculardegeneration (AMD), which is a condition results in a loss of vision inthe center of the visual field (the macula); and (6) neurodegenerativedisease, which is an umbrella term for a range of conditions, whichprimarily affect the neurons in the human brain. Examples ofneurodegenerative diseases contemplated for treatment by the presentinvention include, but are not limited to, Parkinson's Disease (PD),Alzheimer's Disease (AD), Amyotrophic Lateral Sclerosis (ALS or LouGehrig's Disease), Epilepsy, Bi-polar Syndrome, traumatic brain injury,peripheral neuropathy, and Multiple Sclerosis (MS).

BACKGROUND

Malabsorption Syndromes

Malabsorption syndromes encompass a number of different clinicalentities that result in chronic diarrhea, abdominal distention, andfailure to thrive. Clinical malabsorption can be broken down intoseveral distinct conditions, both congenital and acquired, that affectone or more of the different steps in the intestinal hydrolysis andsubsequent transport of nutrients.

Malabsorption is caused by a disorder in the intestinal processes ofdigestion and/or transport of nutrients across the intestinal mucosainto the systemic circulation. Either a congenital abnormality in thedigestive or absorptive processes or, more commonly, a secondarilyacquired disorder of such processes may result in malabsorption.

Neonates, young infants and adolescent humans with malabsorptionsyndromes, in particular fat malabsorption syndromes, are atparticularly high risk for chronic diarrhea and malnutrition due toinsufficient energy intake of high-energy value dietary lipids andessential lipids, which constitute building blocks for normal functionof multiple body systems.

In some humans, exocrine pancreatic insufficiency is the principalcondition that results in severe fat malabsorption. Pancreatitis,pancreatic cancer, pancreatic resection, cystic fibrosis,Shwachman-Diamond syndrome, Johnson-Blizzard syndrome, and Pearsonsyndrome can all result in pancreatic insufficiency. Significantobstructive biliary or cholestatic liver disease or extensive intestinalmucosal disease, such as occurs in celiac disease, may also result insevere steatorrhea.

One particular malabsorption syndrome is Short-Bowel Syndrome (SBS)—adisorder clinically defined by a failure to properly absorb nutrients(malabsorption), frequently accompanied by diarrhea, steatorrhea, fluidand electrolyte disturbances, edema, dehydration and malnutrition.Short-bowel syndrome has numerous causes, both congenital and acquired,but a common etiologic factor is the functional or anatomic loss ofextensive segments of small intestine leading to a severe decrease inintestinal absorptive capacity. Adult human intestines have a normallength range of 260-88 cm. If disease, accident, or any other causeresults in a loss of 50% of the small intestine or leaves less than 200cm of viable length, a patient is at high risk of SBS.

The link between small intestine loss and short-bowel syndrome issimple, losing large amounts of the small intestine compromises thedigestive and absorptive processes. Adequate digestion and absorptioncannot take place and proper nutritional status cannot be maintainedwithout supportive care, frequently including short term and long-termparenteral nutrition. In adults, SBS is commonly caused by radiationenteritis, mesenteric vascular accidents, trauma, and recurrentintestinal obstruction. In children, the most common causes arenecrotizing enterocolitis, intestinal atresias, and intestinal volvulus.Other causes of SBS include congenital short small bowel, gastroschisis,and meconium peritonitis.

Whether or not a patient who has lost a significant amount of smallintestine will develop SBS depends on a number of factors. Importantcofactors that help to determine whether the syndrome will develop ornot include the premorbid length of the small intestine, how muchintestine is lost, the age of the patient, the remaining length of smallintestine and colon, the functional quality of the residual bowel, andthe presence or absence of the ileocecal valve.

Treatment of malabsorption syndromes, such as SBS, is mainly aimed atsupplying the nutrients and vitamins that patients lack. It may include(a) a high-calorie diet that includes vitamins, minerals, carbohydrates,proteins, and fats; (b) injections of vitamins and minerals; (c)administration of drugs that slow down the normal movement of the smallintestine; and (d) feeding through a vein (Total Parenteral Nutrition(TPN)).

Most SBS patients are initially fed with TPN. In many patients,intestinal adaptation, alone or in combination with dietarymodifications, allows weaning from TPN. Unfortunately, many patientscannot be weaned from TPN.

Patients on long-term TPN frequently experience serious metaboliccomplications. The most common complications are hepatic and biliarydisorders manifested by steatosis, fibrosis and cholestasis. Thesedisorders can progress to fulminant liver failure. Prior to 2006,advanced liver disease was the most common cause of death in SBSpatients. In infants with SBS, TPN-associated liver disease wasestimated to occur in 40-60% of those receiving long-term TPN. This islargely due to the fact that infants are currently administered anintravenous solution of DHA, leading to liver injury, Mortality ininfants who develop cholestasis is estimated to be as high as 80%. Inaccordance with the present invention, it is possible to significantlydecrease this treatment-related morbidity by administering substantiallypure DHA in combination with at least one surface active agent, asdescribed herein, effective to spontaneously form a plurality of stablemicelles having a particle size within a range of about 1 μm to about 10μm upon contact with an aqueous medium, or alternatively, byadministering pre-formed micelles produced therefrom, because the DHAavoids the first pass effect, the fatty acid is in the less toxicchylomicron form in blood, and there is no IV fistula issue.

Patients with malabsorption syndromes require products that safely andeffectively deliver all essential nutrients. Many considerTPN-associated liver disease (PNALD) to be caused in part by theintravenous lipid emulsions used to provide fat calories in the PNformulation, although the etiology is unclear. It has been suggestedthat PNALD may be caused by inflammation that is related to the type ofintravenous lipid emulsion used. These fat emulsions cannot beeliminated from PN because to do so would lead to the development ofessential fatty acid deficiency and its concomitant complications.Recent data has shown that the replacement of omega-6 soybean oil lipidemulsions with those comprised primarily of omega-3 fatty acids may leadto reduction or avoidance of PNALD without predisposing the patient toessential fatty acid deficiency.

Given the limited treatment options currently available to patients withfat malabsorption syndromes and the associated side effects of thesetreatments, there is a need for alternative medications that can treatfat malabsorption syndromes. Ideally, such medications should providethe necessary and the essential fatty acids and fat calories for propergrowth, development and maintenance of bodily functions in humans of allages, but especially in neonates, and reduce or even avoid PNALD.

Primary Sclerosing Cholangitis

Primary sclerosing cholangitis (PSC) is a chronic cholestatic liverdisease characterized by progressive inflammatory and fibroticdestruction of the intrahepatic and/or extrahepatic bile ducts. As thebile ducts become inflamed and narrow, bile backs into the liver causingliver cells to become inflamed. Over time this inflammation decreasesblood flow within the liver, increasing pressure in the portal vein. Asthe disease progresses, liver cells die and are replaced with scartissue.

The etiology of PSC is unknown, although, the association of PSC withinflammatory bowel disease, autoimmune diseases, and a host of otherhumoral and cellular immune abnormalities make an immunopathogenicmechanism likely.

Although the initial diagnosis of PSC is usually made within the fourthdecade of life with a mean age of 40 years of age in men and 45 years inwomen, PSC is increasingly diagnosed in infants (in neonates, PSC isreferred to as Neonatal Sclerosing Cholangitis (NSC)), children, andadolescents. PSC predominantly affects males. The diagnosis is based oncharacteristic findings in combination with clinical, biochemical andhistological features. Many patients are asymptomatic at presentation,but symptoms develop over time. Symptoms include itch, lethargy,steatorrhea and vitamin deficiencies, metabolic bone disease, bleedingperistomal varices, bacterial cholangitis, dominant biliary strictures,and in advanced cases cholangiocarcinoma.

There is increasing evidence of a genetic predisposition to PSC, but theexact mechanism of susceptibility has not been well defined.

Immunosuppressants, chelators, and steroids are frequently used to helpcontrol the disease process, but have not shown any significant benefit.Ursodeoxycholic acid has been studied and may improve the liver functionprofile, but a high number of adverse events are associated with thistreatment and there is debate about its effectiveness.

Genotype and phenotype analyses have shown that there is an increasedprevalence of cystic fibrosis transmembrane conductance regulatorprotein (CFTR) abnormalities in adults with PSC. The CFTR proteinfunctions as a channel for the movement of ions in and out of theepithelial cells lining exocrine glands. CFTR dysfunction is associatedwith (1) decreased chloride secretions into the bile canaliculi withsubsequent decrease in osmotic extrusion of water into the lumen,resulting in hyperconcentration and acidification of bile leading toobstruction of intrahepatic bile ductules, secondary inflammation andeventual focal biliary cirrhosis; (2) innate immune defects and leads toan excessive host inflammatory response; (3) fatty acid alterations,specifically a decrease in docosahexaenoic acid (DHA) and an increase inarachidonic acid (AA). As CFTR dysfunction seems to be a contributingfactor in the development of primary sclerosing cholangitis, targetingthe correction of the fatty acid abnormalities and associated defects inimmune defense and inflammatory responses seems to be a promising pathin the treatment of PSC.

Omega-3 fatty acids are known to have anti-inflammatory effects. DHA inparticular is known to be an important regulator of inflammation. It hasbeen recently discovered that resolvins and protectins, two types oflipid mediators derived from DHA, modulate the inflammatory response byactively promoting the resolution of inflammation. DHA has been shown tocorrect fatty acid abnormalities and reverse the development of bileduct injury in CFTR^(−/−) knockout mice. Recent data from a 12-month,open-label pilot study (n=23) to evaluate the safety and effectivenessof DHA for the treatment of PSC showed an increase in serum DHA levelsand a significant decline in alkaline phosphatase in patients with PSC.

Currently there are no effective medical therapies for PSC that resultin significant long-term improvement in outcome. Liver transplantationis the only life-extending therapy for patients with end-stage disease,with all of its associated risks, costs and complications. The mediantime from diagnosis to death or liver transplantation is 9-18 years.While different forms of medical treatment have been tried, there are noestablished, effective therapies that result in significant long-termimprovements in outcome other than liver transplantation. Mosttreatments focus on monitoring liver function, managing symptoms and,when possible, temporarily opening blocked bile ducts. These therapiesdo not target the excessive inflammation and subsequent biliary tractfibrosis associated with the disease. Thus, there is clearly an unmetneed for therapies, especially those that use omega-3 fatty acids, thatcan effectively target the inflammation and fibrosis associated with PSCand that therefore may prevent or limit disease progression.

Non-Alcoholic Fatty Liver Disease and Non-Alcoholic Steatohepatitis

Non-Alcoholic Fatty Liver Disease (NAFLD) is a common, often “silent”liver disease and resembles alcoholic liver disease, but occurs inpeople who drink little or no alcohol. The major feature of NAFLD isexcess fat in the liver, along with inflammation and damage. Most peoplewith this disease feel well and are not aware that they have a liverproblem. Nevertheless, the condition can be severe and can lead tocirrhosis, in which the liver is permanently damaged and scarred and nolonger able to function properly.

Non-alcoholic fatty liver disease (NAFLD) is characterized by increasedhepatic fat accumulation in individuals not consuming excessive alcoholand represents a spectrum of disease ranging from ‘simple’ steatosis tonon-alcoholic steatohepatitis. NAFLD is associated with the metabolicsyndrome and is defined by the presence of ≥5% hepatic steatosis. Riskfactors for the development of NAFLD include central obesity, type IIdiabetes, dyslipidemia, and hypertension. In Western populations, theprevalence of NAFLD may exceed 30%, and can be as high as 88% in theobese. NAFLD is the most common liver disease in children. Given theincreasing prevalence and incidence of these conditions, the globalburden of NAFLD is expected to increase.

Currently, NASH and NAFLD are under diagnosed due to poor diseaseawareness, the insufficiency of non-invasive diagnostic tools and thelack of effective approved therapies. As a confirmed diagnosis of NASHcurrently requires a liver biopsy, patients are often diagnosed after ablood test demonstrating elevated levels of liver enzymes, alanineaminotransferase (ALT), and aspartate aminotransferase (AST).

The primary treatment for NAFLD is weight loss by lifestyle therapyinvolving diet and exercise. Weight loss has been shown to improve liverenzymes, decrease plasma triglycerides and improve liver fatness.Bariatric surgery is an extreme option for reducing weight and theresulting improvements in liver pathology. However, bariatric surgery isnot feasible for the large number of patients having this disease.

Treatments with insulin sensitizers, hypolipidemics and vitamin E havebeen shown to be ineffective due to poor patient compliance, associatedweight gain, and side-effects. Although current evidence from availablerandomized control trials suggests that while thiazolidinediones andvitamin E are effective in reducing liver fat, there are serious safetyconcerns about long-term use of these agents.

Non-alcoholic steatohepatitis (NASH) is a fibrotic manifestation, whichprogresses from NAFLD. Like NAFLD, NASH is usually a silent disease withfew or no symptoms and occurs in people who drink little or no alcohol.The major feature in NASH is fat in the liver, along with inflammationand damage. Most people with NASH feel well and are not aware that theyhave a liver problem. Nevertheless, NASH can be severe and canpredispose the individual to hepatic fibrosis, cirrhosis, and subsequentend-stage liver disease and hepatocellular carcinoma. Not every personwith NASH develops cirrhosis, but once serious scarring or cirrhosis ispresent, few treatments can halt the progression. A person withcirrhosis experiences fluid retention, muscle wasting, bleeding from theintestines, and liver failure. Liver transplantation is the onlytreatment for advanced cirrhosis with liver failure, and transplantationis increasingly performed in people with NASH. NASH ranks as one of themajor causes of cirrhosis in the United States, behind hepatitis C andalcoholic liver disease.

Despite the rapidly increasing incidence of NASH, there are no therapiescurrently approved for the treatment of this common liver disorder. TheNASH market has a significant unmet need for pharmacological optionsthat are effective and well tolerated. Current options for managingpatients with NASH are suboptimal and primarily rely on changes inlifestyle to reduce weight, off-label pharmacotherapy and bariatricsurgery for weight loss. Weight loss is the first recommendation forNASH patients and is associated with a significant improvement insteatosis and overall severity of NASH. However, such improvement isonly statistically significant when patients are able to reduce morethan 7% of their body weight over a sustained period of 48 weeks, whichoccurs in less than 50% of NASH patients. Products utilized off label inthe management of NASH comorbidities include vitamin E, insulinsensitizers such as metformin and pioglitazone, which are used fordiabetic patients, and anti-hyperlipidemic agents, pentoxifylline, andursodiol. High-dose vitamin E has been shown in a clinical study ofnon-diabetic patients to reduce inflammation but not fibrosis. Vitamin Eis not recommended for NASH patients with type-2 diabetes due to lack ofdata, therefore the use of vitamin E is limited. While other off-labelpharmacotherapies demonstrate inconsistent benefits or are associatedwith significant side effects. Bariatric surgery is believed to impactNASH through dramatic weight loss, but it has significant complicationsand drawbacks. These include a host of perioperative risk factors, theneed to adhere to post-surgical diet and nutritional regimens and highcosts. A relatively small number of these procedures are performedannually on NASH patients compared to the overall NASH population, whichwe believe is due to the complications and drawbacks of bariatricsurgery relative to NASH patient numbers. We believe widespreadincreased adoption of bariatric surgery for NASH is impractical based oncost and the large number of patients who would require it. In addition,some retrospective and prospective studies have indicated that theprocedure may worsen fibrosis. Liver transplant is a last resort forlife-threatening complications progressing from NASH. NASH is currentlythe third most common reason for liver transplants in United States andis projected to surpass alcohol-based cirrhosis and viral hepatitis tobecome the leading indication for liver transplant by 2020. Theavailability of liver donors is extremely limited and the cost of aliver transplant is a significant economic burden, with an estimatedcost per procedure of approximately $577,000. Studies have demonstratedthat approximately 22% of patients do not survive the five-year periodpost-transplant. Therefore, a significant unmet need exists foralternative treatments for NAFLD and NASH.

While NAFLD occurs in people of all ages, NASH most often occurs inpeople who are middle aged and overweight or obese. Affected individualsmay also have elevated levels of blood lipids (such as cholesterol andtriglycerides) and many have diabetes. NASH affects 2 to 5 percent ofAmericans. Both NASH and NAFLD are becoming more common, possiblybecause of the greater number of Americans with obesity. In the past 10years, the rate of obesity has doubled in adults and tripled inchildren. Obesity also contributes to diabetes and high bloodcholesterol, which can further complicate the health of a patient withNASH.

There is no current approved treatment for NASH or NAFLD. Accordingly,medications for the treatment of NAFLD and/or NASH represent asubstantial unmet need.

Sickle Cell Diseases

Sickle cell disease, also known as sickle-cell anaemia (SCA) anddrepanocytosis, is a group of hereditary blood disorders that affectshemoglobin, the molecule that delivers oxygen throughout the body viared blood cells. Sickle cell disease is caused by mutations in the HbAgene and is inherited in an autosomal recessive pattern (a single pointmutation in the sixth codon of the β-globin gene). There are four commongenotypes associated with SCD—hemoglobin SS (HbSS), HbSbeta0-thalassemia, hemoglobin SC (HbSC), and HbS beta+-thalassemia.

In individuals with two mutated genes, the resultant abnormal hemoglobinS polymerizes under low oxygen tension and causes abnormal red bloodcells. Because of the polymerization, these cells frequently distortinto a rigid sickle, or crescent, shape, in contrast to disc-shapednormal red blood cells. A person with a single abnormal copy does notexperience symptoms and is said to have sickle-cell trait. A person withsickle cell trait inherits one normal allele and one abnormal alleleencoding hemoglobin S (hemoglobin genotype AS). Sickle cell trait isgenerally regarded as a benign condition, although certain individualswith sickle cell trait may develop complications.

Sickle cell disease is a serious disease that significantly compromisesthe quality of patients' lives and reduces life expectancysignificantly. Signs and symptoms of sickle cell disease usually beginin early childhood. The severity of symptoms varies from person toperson and it has been postulated that clinical manifestations resultfrom complex combinations of genetic, cellular and environmentalfactors. Some people have mild symptoms, while others are frequentlyhospitalized for more serious complications.

Sickle cells have a shorter lifespan than normal red blood cells. Sicklecells also deliver less oxygen to the body's tissues. As a result,patents with sickle cell disease frequently develop anemia. Sickle cellanemia is a common manifestation of sickle cell disease. Patients withanemia experience fatigue, weakness, shortness of breath, dizziness,headaches, and coldness in the hands and feet. Anemia can also causedelayed growth and development in children. The rapid breakdown of redblood cells may also cause jaundice.

Sickle cells are rigid and prone to adhesive interactions with eachother, leukocytes, platelets, plasma and vessel walls. These adhesiveinteractions lead to vaso-occlusion in small blood vessels, with thesickle cells either intact or in pieces. Vaso-occlusion cuts down evenmore on the amount of oxygen flowing to body tissues. Acutevaso-occlusive events are known as crises. Crises can last from hours todays. Some patients have one episode every few years while others havemany episodes each year. Crises can be severe enough to requirehospitalization and can be fatal. Vaso-occlusive events in sickle celldisease are believed to be influenced by multiple factors, includingleucocyte adhesion molecules, inflammatory factors, endothelial cellinteractions, haemorheology and erythrocyte heterogeneity.

Three serious symptoms often accompany crises; pain, organ damage andinfection. It is common in the relevant literature for researchers toalso refer to “pain crisis” and “hemolytic (or “haemolytic”) crisis”(rapidly evolving anemia, leukocytosis, jaundice and fever) and “fattyacid membrane abnormality” fatty as meaningful clinical endpoints.

Deprivation of oxygen-rich blood is especially deleterious to the lungs,kidneys, spleen, and brain. A particularly serious complication ofsickle cell disease is pulmonary hypertension linked to blockages in theblood vessels that supply the lungs. Pulmonary hypertension occurs inabout one-third of adults with sickle cell disease and can lead to heartfailure.

Other serious consequences of the blocked blood vessels are strokes,ulcers of the lower extremities, impaired vision and priapism. Blockageof the blood vessels supplying the spleen may lead to failure of thatorgan, which results in serious infectious conditions such asosteomyelitis, cholecystitis, pneumonia and urinary tract infection.

As a result of the vaso-occlusion and organ damage, sickle cell patientsare often in a near-continuous state of inflammation. They have elevatedstates of markers of inflammation such as C-reactive protein, fibrinogenand interleukin. Sickle cell patients also often have near continuousthrombotic activity, low level most of the time but spiking duringcrises.

There are numerous procedures performed to manage complications ofsickle cell disease. These include blood transfusions, dialysis orkidney transplants, gallbladder removal, hip replacements for avascularnecrosis, eye surgeries and wound care for leg ulcers. Bone marrow orstem cell transplants can cure sickle cell anemia, but are not an optionfor most patients because of a lack of well-matched stem cell donors.Ultimately, sickle cell disease causes multi-organ dysfunction and earlydeath in affected individuals. Many succumb to complications of chronicorgan dysfunction and eventual organ failure.

Unfortunately, for decades the root cause of sickle cell disease wasmisunderstood. Prior to very recently, the majority view was that sicklecell disease was merely the result of sickled red blood cells as aphenotypic manifestation of a single genetic point mutation. However,recent studies have shown that sickle cell disease is, in fact, adisease of systemic inflammation, inducing effects not only on the redblood cells, but also other blood cells and the vessels themselves. Infact, recent research has shown that red blood cells do not sickle insickle cell disease patients until after a cascade of events occurswhich leads to an increased stickiness of the blood cells and vessels.

Once the inflammation occurs, the red blood cells sickle, resulting in afeedback loop cascade, which increases the magnitude of theinflammation. This increased inflammation is manifested through theincreased presence of arachidonic acid and a decrease in certain omega-3acids, such as DHA. Thus, without being bound to any particular theory,it is believed that restoring the fatty acid compositions to nearer tonormal levels will create a significant decrease in the inflammation andthus diminish the morbidity of sickle cell disease.

Unless a specific condition, disease state or malady associated withsickle cell disease is referenced herein, as used herein, a reference to“sickle cell disease” or “sickle cell condition” or “sickle cell diseasecondition” refers to each and all of the disease states and/or effectsand/or conditions of sickle cell disease as described herein or asotherwise known by the ordinarily skilled artisan in the relevant art. Areference to “at least one sickle cell condition” refers to one or moreof the conditions related to sickle cell disease as described herein oras otherwise known to such ordinarily skilled artisan.

Existing treatments for sickle cell disease are primarily aimed atrelieving symptoms, limiting the number of crises and lesseningcomplications. People with sickle cell disease need ongoing treatment,even when not having a crisis.

Frequently prescribed therapeutic compositions, thought to be of benefitfor sickle cell patients, include pain medications, antibiotics, folicacid, hydroxyurea, and anti-inflammatory medications. Pain medicines ofall types are heavily prescribed to address the pain episodes associatedwith sickle cell disease. Antibiotics are frequently prescribed to treatthe bacterial infections that are common in sickle cell patients. Sicklecell patients are often advised to take folic acid supplements becausefolic acid aids in the production of new red blood cells.

Hydroxyurea (tradename DROXIA) is approved to reduce the number of painepisodes (including chest pain and breathing problems) and bloodtransfusions in patients with recurring moderate to severe crises but isa cytotoxic molecule with teratogenic and carcinogenic effects. Theprecise mechanism of action of hydroxyurea is uncertain. It has beenfound that besides the damage caused by sickle cells themselves, theinflammatory response that occurs in sickle cell disease patients couldpotentially play a significant role in the occurrence of painfulepisodes or pain crises. Sickle cell patients experience relief ofsymptoms when treated with a variety of anti-inflammatory agents.

As early as 1991, it was suggested that omega-3 fatty acids decrease thehemolysis of mammalian red blood cells. It also has been found thatsickle cell patients have abnormal blood fatty acids. These findingshave led to the hypothesis that omega-3 fatty acids may be useful in thetreatment of sickle cell disease. As early as 2001, small human clinicaltrials showed that omega-3 fatty acids could reduce pain episodes insickle cell patients, perhaps by reducing prothrombotic activity. Otherstudies have shown that omega-3 fatty acids can increase hemoglobinlevels, and reduce pain episodes, vaso-occlusive episodes, anemia, organdamage and other disease complications in sickle cell patients.

Clupanodonic acid, also known as omega-3 DPA, is known to be useful as aplatelet aggregation inhibitor and other omega-3 acids are known topossess anti-inflammatory properties. Vaso-occlusive events are thesource of a number of problematic symptoms in patients with sickledisease and can occur through platelet aggregation as well as red bloodcell aggregation and white blood cell aggregation. Some vaso-occlusiveevents are the result of thrombosis, which is initiated by thromboxaneA2 (TXA2). Omega-3 DPA has been shown to be a potent inhibitor of COX-1activity (the enzyme involved in synthesis of TXA2), thus omega-3 DPAcan inhibit the pathways through which vaso-occlusive events occur. Thestudies above strongly suggest that omega-3 fatty acids can delay oreven prevent some of the damaging effects of inflammation in sickle cellpatients. Given the lack of effective drugs in treating sickle celldisease, there is a critical unmet need in developing new therapies forcombatting this debilitating and life-shortening disease.

Accordingly, any new therapy that reduces the frequency or severity ofinflammation, anemia and/or vaso-occlusive crises in patients is likelyto play a useful role in the treatment of sickle cell disease. Given theobservation that omega-3 fatty acids may be useful in the treatment ofsickle cell disease and improving or protecting cell health, newformulations, particularly those with a demonstrable increase inabsorption and blood levels of omega-3 and other fatty acids in thetreatment of sickle cell disease are warranted.

Macular Degeneration

Age-related macular degeneration (AMD) is a condition that results in aloss of vision in the center of the visual field (the macula). It is aprogressive disease and is the leading cause of severe vision loss inpeople over age 60. It occurs in “dry” and “wet” forms and causes damageto the macula, a small spot near the center of the retina and the partof the eye needed for sharp, central vision, which lets us see objectsthat are straight ahead.

The macula is the part of the retina that receives most of the directlight in the eye; is very sensitive to light, with higher concentrationsof rods and cones, and is needed for accurate central vision. The retinais a network of visual receptors and nerves that lies on the choroid; alayer of blood vessels that provides blood to the macula, and theretinal pigment epithelium (RPE). The RPE is another layer ofspecialized cells, which transports the nutrients from the blood vesselsand clears waste products from the macula.

Dry AMD is an early stage of the disease and may result from the agingand thinning of macular tissues due to the degeneration or atrophy thatoccurs from cell death. It is characterized by the presence of yellowspots, called drusen, which form in and around the macula. It isbelieved these spots are deposits or debris from deteriorating tissue,which accumulates between the retina and the choroid due to theinability of the RPE to get rid of the waste and leads to the eventualdeterioration of the photoreceptors.

Wet AMD refers to the growth of abnormal growth of blood vessels(angiogenesis) beneath the retina, also referred to as choroidalneovascularization. These blood vessels leak blood and fluid into theretina, causing distortion of vision that makes straight lines lookwavy, as well as blind spots and loss of central vision. These abnormalblood vessels eventually scar, leading to permanent loss of centralvision.

Dry AMD occurs less often to people who exercise, avoid smoking and eathealthy food, including vegetables and fish. The National Eye Institutefound in the Age-Related Eye Disease Studies (AREDS) that daily intakeof a combination of vitamin C, vitamin E, beta-carotene, zinc, andcopper can reduce the risk of late AMD by 25 percent and can slowprogression of the disease. The AREDS2 trial found that adding luteinand zeaxanthin or omega-three fatty acids to the original AREDSformulation (with beta-carotene) had no overall effect on the risk oflate AMD. However, the trial also found that replacing beta-carotenewith a 5-to-1 mixture of lutein and zeaxanthin may help further reducethe risk of late AMD. While lutein and zeaxanthin appear to be saferegardless of smoking status of the individual suffering from AMD, highbeta-carotene intake has been linked to an increased risk of lung cancerin current and former smokers. For wet AMD, angiogenesis inhibitors,such as for example, BEVACIZUMAB, RANIBIZUMAB, PEGAPTANIB, ANDAFLIBERCEPT can be injected into one's eye to stop new blood vesselsfrom growing.

Oxidative damage, resulting from excess production of reactive oxygenspecies (ROS) has been implicated in the progression AMD. Retinalpigmented epithelial (RPE) cells are highly metabolically active andthere is strong evidence that the RPE cells are extremely sensitive tooxidative stress. It has been reported that the pathophysiology of AMDis due to cumulative oxidative damage to RPE cells resulting from animbalance between the generation of ROS and the ability of these cellsto destroy and/or protect against ROS damage. Strategies for protectingRPE cells against oxidative damage may be particularly important inmaintaining retinal function and both preventing and treating AMD.

AREDS was designed to evaluate the prevention and treatment effects ofhigh doses of antioxidants and zinc on individuals with AMD. AREDS2, wasconducted to evaluate the effects on progression and treatment of AMDpatients of a modified version of the AREDS formulation, includingomega-3 fatty acids. The study showed no AMD-related benefit from theintroduction of the omega-3 fatty acids into the treatment regimen.However, because the AREDS2 formulation did not include formulas withself-micellizing properties, the AREDS2 study was unable to evaluate anefficacious bioavailability of the omega-3 fatty acids on AMD.Furthermore, the AREDS2 study was unable to evaluate predictable,non-food dependent bioavailability of fat-soluble vitamins because saidvitamins were not co-administered with self-micellizing formulations.Therefore, the benefits of administering omega-3 fatty acids withself-micellizing formulations would not have been recognized by previousstudies.

The lack of any approved effective medications for AMD warrants thecritical need for new therapies to treat AMD.

Neurodegenerative Diseases

Neurodegenerative disease is an umbrella term for a range of conditions,which primarily affect the neurons in the human brain. Examples of themore common neurodegenerative diseases include Parkinson's Disease (PD),Alzheimer's Disease (AD), Amyotrophic Lateral Sclerosis (ALS or LouGehrig's Disease), Epilepsy, and Multiple Sclerosis (MS).

PD is a neurodegenerative disorder primarily characterized by motorsymptoms but which also includes several other pathological featuressuch as autonomic system failures, mood disorders, and cognitivedeficits. Conventional treatment may involve medication that isprimarily aimed at increasing dopamine activity either by providing theprecursor (raw material) in the form of levodopa (L-Dopa), or bystimulating dopamine receptors (essentially mimicking dopamine) throughthe use of a dopamine agonist drug. Also used are drugs called COMTinhibitors which can help the levodopa to be more effective and MAO-Binhibitors which prevent dopamine from breaking down so the limitedsupply is longer lasting. Other therapies may include physiotherapy,osteopathy, remedial movement, massage, speech therapy, psychologicaltherapy and in some cases surgery (deep brain stimulation). Currentpharmacological options for the disease are limited to symptommanagement and their long-term use leads to important side effects.

MS is a nervous system disease that affects your brain and spinal cord.It damages the myelin sheath, the material that surrounds and protectsyour nerve cells. This damage slows down or blocks messages between yourbrain and your body, leading to the symptoms of MS. They can includevisual disturbances, muscle weakness, trouble with coordination andbalance, sensations such as numbness, prickling, or “pins and needles”,and thinking and memory problems. Although there is no known cure formultiple sclerosis, several therapies have proven helpful. The primaryaims of therapy are returning function after an attack, preventing newattacks, and preventing disability. As with any medical treatment,medications used in the management of MS have several adverse effects.Available management and/or treatment of the disease is dependent on thestage of the disease. For example, in relapsing remitting multiplesclerosis (RRMS), eight disease-modifying treatments have been approvedby regulatory agencies for RRMS including: interferon beta-la,interferon beta-1b, glatiramer acetate, mitoxantrone, natalizumab,fingolimod, teriflunomide, and dimethyl fumarate. There is no treatmentavailable, however, for primary progressive MS (PPMS).

ALS, is a rapidly progressive, neuromuscular disease. It attacks themotor neurons that transmit electrical impulses from the brain to thevoluntary muscles in the body. When they fail to receive messages, themuscles lose strength, atrophy and die. RILUZOLE, which extends survivalof ALS patients by several months, is currently the only treatmentapproved for improving survival, but does not reverse the damage alreadydone to motor neurons. BACLOFEN and DIAZEPAM are often prescribed tocontrol the spasticity caused by ALS, and trihexyphenidyl oramitriptyline may be prescribed when ALS patients begin having troubleswallowing their saliva.

AD is an irreversible, progressive brain disease that slowly destroysmemory and thinking skills, and eventually even the ability to carry outthe simplest tasks. AD is the most common cause of dementia among olderpeople. Dementia is the loss of cognitive functioning—thinking,remembering, and reasoning—and behavioral abilities, to such an extentthat it interferes with a person's daily life and activities. Dementiaranges in severity from the mildest stage, when it is just beginning toaffect a person's functioning, to the most severe stage, when the personmust depend completely on others for basic activities of daily living.Four medications are approved by the U.S. Food and Drug Administrationto treat Alzheimer's. DONEPEZIL (ARICEPT®), RIVASTIGMINE (EXELON®), ANDGALANTAMINE (RAZADYNE®) are used to treat mild to moderate Alzheimer's(donepezil can be used for severe Alzheimer's as well). MEMANTINE(NAMENDA®) is used to treat moderate to severe Alzheimer's. These drugswork by regulating neurotransmitters (the chemicals that transmitmessages between neurons). They may help maintain thinking, memory, andspeaking skills, and help with certain behavioral problems. However,these drugs don't change the underlying disease process, are effectivefor some but not all people, and may help only for a limited time.

Epilepsy is a group of neurological diseases characterized by epilepticseizures. Epileptic seizures are episodes that can vary from brief andnearly undetectable to long periods of vigorous shaking. In epilepsy,seizures tend to recur, and have no immediate underlying cause whileseizures that occur due to a specific cause are not deemed to representepilepsy. The cause of most cases of epilepsy is unknown, although somepeople develop epilepsy as the result of brain injury, stroke, braintumor, and substance use disorders. Genetic mutations are linked to asmall proportion of the disease. Epileptic seizures are believed to bethe result of excessive and abnormal cortical nerve cell activity in thebrain. Seizures are controllable with medication in about 70% of cases.In those whose seizures do not respond to medication, then surgery,neurostimulation, or dietary changes may be considered. The mainstaytreatment of epilepsy is anticonvulsant medications, a number of whichare available. Phenobarbital, phenytoin, carbamazepine and valproateappear to be equally effective in both partial and generalized seizures.

Current epidemiological, preclinical and clinical data suggest thatomega-3 polyunsaturated fatty acids (n-3 PUFAs) may constitutetherapeutic strategy for several disorders of the central nervoussystem, including PD, AD, ALS, Epilepsy, Bi-polar Syndrome, traumaticbrain injury, peripheral neuropathy, and MS. These fatty acids, foundmost commonly in certain fish and some plants, are known to help reduceinflammation and oxidative stress on cells. Both of these processes areknown to damage nerve tissue.

For example, the concentration of omega-3 fatty acids in nerve cellmembranes has been shown to decrease with age, oxidant stress, and inneurodegenerative disorders such as Parkinson's disease. Researchers inNorway have presented evidence of a systematic omega-3 fatty aciddeficit in PD and AD suggesting a fundamental neurological role forthese vital fat molecules. Supplementation with the omega-3 DHA canfavorably modify brain functions and has been proposed as anutraceutical in PD and AD. More recently, the organization OvercomingMultiple Sclerosis (OMS) conducted a clinical trial looking for a linkbetween omega-3 fatty acids and MS via the international HOLISM study.The findings strongly supported a link between consumption of fish andomega-3 supplements and better health for people with MS. Specifically,people with MS taking flaxseed oil regularly had over 60% fewer relapsesthan those who didn't in this large international sample. Association offish consumption and omega-3 supplementation with quality of life,disability and disease activity has been studied in an internationalcohort of people with multiple sclerosis.

Similarly, recent studies show consumption of foods high in omega-3fatty acids may help prevent or delay the onset of ALS.

The studies above strongly suggest that omega-3 fatty acids can delay oreven prevent some neurodegenerative diseases. Given the lack ofeffective drugs in treating these diseases, there is a critical unmetneed in developing new therapies for combatting at least certainneurodegenerative diseases.

Nutritional Deficiencies and their Effect on Disease StateManifestations

Studies have demonstrated that vitamin D may play a significant role ineye health, specifically in the prevention of AMD. A high correlation isknown to exist between deficient vitamin D levels and AMD. Vitamin D isbelieved to act through the vitamin's anti-inflammatory oranti-angiogenic properties. For this reason, vitamin D may be useful inthe treatment of multiple inflammation related diseases, including AMD,sickle cell disease, Alzheimer's, and other diseases.

Zinc is one of the most abundant trace metals in the human body. Over300 enzymes are known to utilize zinc in normal mammalian function. Zincdeficiency affects many functions that are directly or indirectlyrelated to declining cognitive performance in aging individuals,including the progression of Alzheimer's. Because zinc use is soprolific throughout the body, zinc deficiency can be causally related toAlzheimer's disease through multiple pathways. For instance, zincdeficiency can induce insulin resistance, a risk factor for Alzheimer's.Furthermore, zinc is believed to be very important in the transport oflipids across the intestinal lining and therefore affect many diseasesrelated to lipid deficiencies, including Alzheimer's.

Alzheimer's disease represents a major health problem in the US,estimated to be the third leading cause of death, yet the causes ofAlzheimer's disease remain largely unknown and misunderstood. However,recent research is beginning to demonstrate a clear link between theprogression of the disease and certain changes in lipids, includingfatty acids and fat soluble vitamins, as well as magnesium and zinc.Markers correlated with progression of Alzheimer's include changingfatty acid profiles of DHA, EPA, and arachidonic acid, as well asmetabolic syndrome, chronic inflammation, hypovitaminosis D, and zincdeficiency. Furthermore, these markers themselves may be correlated.

Treatment of these correlated conditions can be made through theintroduction of lipids in the correct ratios or amounts. However,because the absorption of lipids by the digestive system requires aco-consumption of the dietary fats necessary to form naturally occurringmicelles, the ability to properly dose patients is often a significantchallenge. Further complicating matters, the over-introduction ofcertain lipids, such as the fat-soluble vitamins, can be harmful.Therefore, a need exists to provide the necessary lipids in a mannerwhich is diet-independent such that dosing can be more tightlycontrolled.

Magnesium contributes to more than three hundred chemical reactions inmammals, including every ATP-dependent reaction. Magnesium deficiency ismost commonly associated with malabsorption diseases, including Crohn'sdisease, gluten-sensitive enteropathy (celiac disease), and regionalenteritis. Resection or bypass of the small intestine, such as in shortbowel disease, typically leads to malabsorption and magnesium loss. Theinteraction of magnesium and vitamin D is particularly important acrossa plethora of pathways and body systems. For example, magnesium isrequired for both steps in the activation of vitamin D to calcitriol,the form of the vitamin necessary for calcium absorption. Magnesium isalso required for calcitriol's role in calcium absorption. Mammals withlow magnesium are deficient in both vitamin D and calcitriol, but theintroduction of calcitriol alone does not improve calcium absorption.Magnesium deficits are correlated to type 2 diabetes and insulinresistance as a result of higher excretion of magnesium that resultsfrom increased glucose concentrations in the kidneys. Thus, a needexists to co-administer magnesium with lipids in the treatment orprevention of certain disease states.

Vitamins A, D, and K cooperate synergistically not only with each other,but also with essential minerals like magnesium and zinc, with dietaryfats. Vitamins A, D, and K2 interact synergistically to support immunehealth, support bone and teeth strengthening, and protect soft tissuesfrom calcification. The interaction of these fat-soluble vitamins withmagnesium and zinc is important. Zinc supports the formation of vesiclesinvolved in transporting lipids, including the fat-soluble vitamins,across the intestinal wall.

SUMMARY OF THE INVENTION

Provided herein, in certain embodiments, are self-micellizingcompositions comprising at least one omega-3 fatty acid composition(either in the triglyceride, ester, or free fatty acid form) and atleast one surface active agent composition. These compositions, whencombined in specific ratios, are characterized by their ability tospontaneously form a plurality of stable micelles within a range ofabout 1 μm to about 10 μm upon contact with an aqueous medium.

In order to determine sample size in a uniform and reproducible manner,all samples were characterized utilizing the LS™ 13 320 Aqueous LiquidModule (ALM).

The LS™ 13 320 Laser Diffraction Particle Size Analyzer uses laserdiffraction and a patented polarization intensity differentialscattering (PIDS) technology to rapidly determine the particle sizedistribution of materials with an overall sizing range of 0.04 μm to2000 μm in a single scan with no extrapolation. This is accomplishedwith high resolution and excellent reproducibility.

In accordance with a standardized protocol, the sample was heated in itsoriginal container at 37° C. for one hour. After mixing well,approximately 0.25 g of sample was diluted with 5 mL of deionized water.The dilution was manually shaken to form a homogenous mixture. Thepreparation was allowed to equilibrate to room temperature and wassubsequently dispensed to the ALM sample vessel dropwise and analyzedfor the particle size distribution (PSD), which was determined using theFraunhofer optical model. The instrument analyzed the PSD from 0.040 μmto 2,000 μm.

In certain embodiments, the at least one omega-3 fatty acid (either inthe triglyceride, ester, or free fatty acid form) is selected from thegroup consisting of hexadecatrienoic acid, α-linolenic acid, stearidonicacid, eicosatrienoic acid, eicosapentaenoic acid, heneicosapentaenoicacid, the omega-3 isomer of docosapentaenoic acid, docosahexaenoic acid,tetracosapentaenoic acid, and tetracosahexaenoic acid. In certainembodiments, self-micellizing compositions may further contain anomega-6 fatty acid (either in the triglyceride, ester, or free fattyacid form), for example arachidonic acid (ARA), linoleic acid (LA),Gamma-linolenic acid (GLA), and omega-6 docosapentaenoic acid (omega-6DPA).

Certain embodiments provide for at least one surface active agent, afirst omega-3 fatty acid (either in the triglyceride, ester, or freefatty acid form) and a second omega-3 fatty acid (either in thetriglyceride, ester, or free fatty acid form), wherein the first andsecond omega-3 fatty acids are different and are selected from the groupconsisting of hexadecatrienoic acid, α-linolenic acid, stearidonic acid,eicosatrienoic acid, eicosapentaenoic acid, heneicosapentaenoic acid,the omega-3 isomer of docosapentaenoic acid, docosahexaenoic acid,tetracosapentaenoic acid, and tetracosahexaenoic acid.

Certain embodiments provide for compositions comprising at least onesurface active agent and a first omega-3 fatty acid (either in thetriglyceride, ester or free fatty acid ester form) and a second omega-3fatty acid (in either the ester, triglyceride or free fatty acid form),wherein the first and second omega-3 fatty acids are different andwherein the ratio of the amount of the first omega-3 fatty acid to thesecond omega-3 fatty acid is 1:X where 0<X<1. In certain otherembodiments, X can be, for example, 0.001, 0.0015, 0.002, 0.0025,0.0055, 0.006, 0.0065, 0.007, 0.0075, 0.008, 0.0085, 0.009, 0.0095,0.01, 0.015, 0.02, 0.025, 0.03, 0.035, 0.04, 0.045, 0.05, 0.055, 0.06,0.065, 0.07, 0.075, 0.08, 0.085, 0.09, 0.095, 0.1, 0.15, 0.2, 0.25,0.55, 0.6, 0.65, 0.7, 0.75, 0.8, 0.85, 0.9, or, 0.95.

In certain embodiments, the compositions comprise at least one surfaceactive agent and a first and second omega-3 fatty acid, wherein thefirst omega-3 fatty acid (in either the triglyceride, ester, or freefatty acid form) is docosahexaenoic acid (DHA) and the second omega-3fatty acid (in either the triglyceride, ester, or free fatty acid form)is selected from the group consisting of hexadecatrienoic acid,α-linolenic acid, stearidonic acid, eicosatrienoic acid,eicosapentaenoic acid, heneicosapentaenoic acid, the omega-3 isomer ofdocosapentaenoic acid, tetracosapentaenoic acid, and tetracosahexaenoicacid and combinations thereof.

In certain other embodiments, the compositions comprise at least onesurface active agent and a first and second omega-3 fatty acid, whereinthe first omega-3 fatty acid (in either the triglyceride, ester, or freefatty acid form) is eicosapentaenoic acid (EPA) and the second omega-3fatty acid (in either the triglyceride, ester, or free fatty acid form)is selected from the group consisting of hexadecatrienoic acid,α-linolenic acid, stearidonic acid, eicosatrienoic acid,heneicosapentaenoic acid, docosahexaenoic acid (DHA), the omega-3 isomerof docosapentaenoic acid (DPA), tetracosapentaenoic acid, andtetracosahexaenoic acid and combinations thereof.

In certain embodiments, the compositions comprise at least one surfaceactive agent and a first and second omega-3 fatty acid, wherein thefirst omega-3 fatty acid (in either the triglyceride, ester, or freefatty acid form) is DHA and the second omega-3 fatty acid (in either thetriglyceride, ester, or free fatty acid form) is EPA, wherein the ratioof the amount of DHA:EPA is 1:X, where 0<X<1. In certain otherembodiments, X can be, for example, 0.001, 0.0015, 0.002, 0.0025,0.0055, 0.006, 0.0065, 0.007, 0.0075, 0.008, 0.0085, 0.009, 0.0095,0.01, 0.015, 0.02, 0.025, 0.03, 0.035, 0.04, 0.045, 0.05, 0.055, 0.06,0.065, 0.07, 0.075, 0.08, 0.085, 0.09, 0.095, 0.1, 0.15, 0.2, 0.25,0.55, 0.6, 0.65, 0.7, 0.75, or 0.8.

In certain embodiments, the compositions comprise at least one surfaceactive agent and a first and second omega-3 fatty acid, wherein thefirst omega-3 fatty acid (in either the triglyceride, ester, or freefatty acid form) is EPA and the second omega-3 fatty acid (in either thetriglyceride, ester, or free fatty acid form) is DHA, wherein the ratioof the amount of EPA:DHA is 1:X, where 0<X<1. In certain otherembodiments, X can be, for example, 0.001, 0.0015, 0.002, 0.0025,0.0055, 0.006, 0.0065, 0.007, 0.0075, 0.008, 0.0085, 0.009, 0.0095,0.01, 0.015, 0.02, 0.025, 0.03, 0.035, 0.04, 0.045, 0.05, 0.055, 0.06,0.065, 0.07, 0.075, 0.08, 0.085, 0.09, 0.095, 0.1, 0.15, 0.2, 0.25,0.55, 0.6, 0.65, 0.7, 0.75, 0.8, 0.85, 0.9, or 0.95.

In certain embodiments, the compositions described herein optionallycomprise a third fatty acid (either in the triglyceride, ester, or freefatty acid form), wherein the third fatty acid is different from thefirst and second omega-3 fatty acids. In such compositions, the ratio ofthe amount of the first omega-3 fatty acid to the second and thirdomega-3 fatty acids is 1:X, where X is the combined amount of the secondand third fatty acids and where 0<X<1. For example, if the first omega-3fatty acid is DHA and the second omega-3 fatty acid is EPA, the thirdomega-3 fatty acid may be selected from the group consisting ofhexadecatrienoic acid, α-linolenic acid, stearidonic acid,eicosatrienoic acid, heneicosapentaenoic acid, the omega-3 isomer ofdocosapentaenoic acid, tetracosapentaenoic acid, and tetracosahexaenoicacid. The ratio of the amount of DHA:(EPA+third omega-3 fatty acid) is1:X, where X is the amount of EPA plus the amount of the third omega-3fatty acid, and where 0<X<1. In certain embodiments, X can be, forexample, 0.001, 0.0015, 0.002, 0.0025, 0.0055, 0.006, 0.0065, 0.007,0.0075, 0.008, 0.0085, 0.009, 0.0095, 0.01, 0.015, 0.02, 0.025, 0.03,0.035, 0.04, 0.045, 0.05, 0.055, 0.06, 0.065, 0.07, 0.075, 0.08, 0.085,0.09, 0.095, 0.1, 0.15, 0.2, 0.25, 0.3, 0.35, 0.4, 0.45, 0.5, 0.55, 0.6,0.65, 0.7, 0.75, 0.8, 0.85, 0.9, or 0.95. In certain embodiments, thethird fatty acid can be an omega-6 fatty acid such as arachidonic acid(ARA), linoleic acid (LA), gamma-linolenic acid (GLA) or the omega-6isomer of DPA (either in the triglyceride, ester, or free fatty acidform).

In certain embodiments, the omega-3 fatty acids (either in thetriglyceride, ester, or free fatty acid form) are substantially pure.

In certain embodiments, the omega-3 fatty acids are in the form oftriglycerides.

In certain embodiments, the omega-3 fatty acids are in the form ofesters.

In certain embodiments, the omega-3 fatty acid ester is the ethyl esterderivative.

In certain embodiments, the omega-3 fatty acids are in the form of freefatty acids.

In certain embodiments, the compositions comprising the at least onesurface active agent and the omega-3 fatty acids in either thetriglyceride or ester forms are free of free fatty acid forms.

In certain embodiments, the compositions are substantially free ofactive agents other than the omega-3 fatty acids (either in thetriglyceride, ester, or free fatty acid form).

In certain embodiments, the omega-3 fatty acid or combinations ofomega-3 fatty acids (in either the triglyceride, ester, or free fattyacid form) comprises, for example, 60%, 65%, 70%, 75%, 80%, 85%, 90% or95% (wt/wt) of the composition.

In certain embodiments, the compositions described herein comprise DHAat, for example, from 50% to 99% (wt/wt) of the compositions. Forexample, in certain embodiments, the DHA is present at 50%, 55%, 60%,65%, 70%, 75%, 80%, 85%, 90%, 95%, or 99% (wt/wt) of the compositions.In certain other embodiments, the DHA is present at least, for example,90% (wt/wt) of the compositions described herein. In still otherembodiments, the DHA is present at least, for example, 95% (wt/wt) ofthe compositions described herein.

In certain embodiments, the compositions described herein comprise EPAat, for example, from 0% to 50% (wt/wt) of the compositions. Forexample, in certain embodiments, the EPA is present at <1% (wt/wt) ofsaid compositions. In certain other embodiments, the EPA can be presentat, for example, 2%, 4%, 6%, 8%, 10%, 12%, 14%, 16%, 18%, 20%, 22%, 24%,26%, 28%, 30%, 32%, 34%, 36%, 38%, 40%, 42%, 44%, 46%, 48% or 50%(wt/wt) of said compositions. In still other embodiments, the EPA can bepresent at <5% (wt/wt) of said compositions. For example, in certainembodiments, the EPA can be present at 0%, 0.1%, 0.2%, 0.3%, 0.4%, 0.5%,0.6%, 0.7%, 0.8%, 0.9%, 1%, 1.5%, 2%, 2.5%, 3%, 3.5%, 4%, or 4.5%(wt/wt) of said compositions.

In certain embodiments, the compositions described herein comprise anomega-3 fatty acid other than DHA and EPA at, for example, from >0% to10% (wt/wt) of said compositions. For example, said omega-3 fatty acidother than DHA and EPA can be present, in certain embodiments at 0.1%,0.2%, 0.3%, 0.4%, 0.5%, 0.6%, 0.7%, 0.8%, 0.9%, 1%, 1.5%, 2%, 2.5%, 3%,3.5%, 4%, 4.5%, 5%, 5.5%, %, 6.5%, 7%, 7.5%, 8%, 8.5%, 9%, 9.5% or 10%(wt/wt) of said compositions. In such embodiments, the omega-3 fattyacid other than EPA and DHA can be selected from the group consisting ofhexadecatrienoic acid, α-linolenic acid, stearidonic acid,eicosatrienoic acid, heneicosapentaenoic acid, the omega-3 isomer ofdocosapentaenoic acid, tetracosapentaenoic acid, and tetracosahexaenoicacid.

In certain embodiments, the compositions described herein comprise 90%,5%, and 5% (wt/wt) of said compositions DHA, ARA, and EPA respectively.

In certain embodiments, the compositions described herein comprise 95%,5%, and <1% (wt/wt) of said compositions DHA, ARA, and EPA respectively.

In certain embodiments, the omega-3 fatty acid compositions describedherein comprise at least one surface active agent selected from thegroup consisting of at least one nonionic surface active agents,cationic surface active agents, anionic surface active agents,zwitterionic surface active agents, and combinations thereof.

In certain embodiments, the surface active agent is selected from thegroup consisting of at least one anionic surface active agent, at leastone non-ionic surface active agent, and a combination thereof.

In certain embodiments comprising at least one surface active agent, theat least one surface active agent has a hydrophilic-lipophilic balance(HLB) of 8.0.

In certain embodiments comprising at least one surface active agent, thesurface active agent can be a non-ionic surface active agent selectedfrom the group consisting of at least one polysorbate, at least onepoloxamer, and a combination thereof.

In certain embodiments, the at least one surface active agent comprisesa polysorbate present from, for example, 15% (wt/wt) to 35% (wt/wt) ofthe composition.

In certain embodiments, the polysorbate is present at, for example, 31%(wt/wt) of the composition.

In certain embodiments, the polysorbate is polysorbate 80.

In certain other embodiments, the at least one surface active agentcomprises a poloxamer present from, for example, 0.1% (wt/wt) to 5%(wt/wt) of the composition.

In certain embodiments, the poloxamer is present at, for example, 0.7%(wt/wt) of the composition.

In certain embodiments, the poloxamer is Poloxamer 237, also known asPluronic® F87.

In certain embodiments, the compositions described herein comprise acombination of polysorbate 80 and the Poloxamer 237 (Pluronic® F87)[(HO(C₂H₄O)₆₄(C₃H₆O)₃₇(C₂H₄O)₆₄H].

In certain embodiments, the combination of polysorbate and poloxamercomprise at least, for example, 25% (wt/wt) of said composition.

In certain embodiments, the composition further comprises at least oneantioxidant. In such embodiments the at least one antioxidant isselected from the group consisting of a tocopherol, a tocotrienol, andcombinations thereof. In such embodiments, the tocopherol, tocotrienoland combinations thereof is present from, for example, from about 0.01%to about 5% by weight of the compositions. In certain such embodiments,the tocopherols, tocotrienols and combinations thereof can be presentat, for example, 0.01%, 0.05%, 0.1%, 0.2%, 0.3%, 0.4%, 0.5%, 0.6%, 0.7%,0.8%, 0.9%, 1%, 1.5%, 2%, 2.5%, 3%, 3.5%, 4%, 4.5%, 5%, 5.5%, or 6%(wt/wt) of the compositions. In certain such embodiments, thetocopherols, tocotrienols, and combinations thereof can be present at,for example, 1%, 1.5%, 2%, 2.5%, 3%, 3.5%, 4%, 4.5%, 5%, 5.5%, or 6%(wt/wt) of the compositions. In certain embodiments further comprisingat least one antioxidant, the antioxidant is d-gamma tocotrienol presentat, for example, 3%, 3.35%, 4%, 4.35%, 5%, or 5.35% (wt/wt) of thecomposition. In certain embodiments further comprising at least oneantioxidant, the antioxidant is α-tocopherol present at, for example, 2%(wt/wt) of the composition.

Certain embodiments provide for compositions comprising at least oneterpene. Said present can be present from, for example, 0.1% (wt/wt) to5% (wt/wt). In certain embodiments, the terpene is present at, forexample, 0.01%, 0.05%, 0.1%, 0.2%, 0.3%, 0.4%, 0.5%, 0.6%, 0.7%, 0.8%,0.9%, 1%, 1.5%, 2%, 2.5%, 3%, 3.5%, 4%, 4.5% or 5% (wt/wt) of saidcomposition. The terpene can be d-limonene.

In embodiments comprising substantially pure d-limonene, the d-limoneneis from about, for example, 95% to about 98% pure. In certainembodiments, the substantially pure d-limonene is at least, for example,95%, 96%, 97% or 98%, 99% pure.

Certain embodiments provide for compositions comprising naturalorange-oil.

In certain embodiments, the bioavailability of the omega-3 fatty acids(in either the triglyceride, ester, or free fatty acid form) included inthe compositions described herein is substantially the same whenadministered with or without food, i.e., substantially independent offood effect.

While not wishing to be bound to any particular theories, it has beenobserved that compositions comprising at least one omega-3 fatty acid(either in the triglyceride, ester or free fatty acid ester form) and atleast one surface active agent, which are present in an amount and acombination effective to cause said compositions to self-micellize whenin contact with an aqueous medium, can be characterized by their abilityto spontaneously form a plurality of stable micelles having a particlesize within a range of about 1 μm to about 10 μm, upon contact with theaqueous medium. The plurality of micelles produced in this mannerdemonstrate enhanced bioavailability of said at least one omega-3 fattyacid, while eliminating any food effect, and improving patientcompliance due to the lowered dosage required to reach a similarbioavailability or AUC (e.g. 1 pill versus 6).

In at least one embodiment, a method is provided for treating AMD byadministering the compositions described herein to a human in need ofsuch administration. The human can be an adult, a child, an adolescent,or and infant, such as for example a neonate. In certain embodiments, amethod is provided for treating Juvenile Macular Degeneration, whichincludes Stargardt's disease, Best disease, and juvenile retinoschisisby administering the compositions described herein to a human juvenile.

In certain embodiments, the compositions may comprise at least oneomega-3 fatty acid, and at least one non-omega-3 fatty acid nutritionalsupplement agent, wherein the omega-3 fatty acid (in either thetriglyceride, ester, or free fatty acid form) is docosahexaenoic acid(DHA) (in either the triglyceride, ester, or free fatty acid form) andwherein the non-omega-3 fatty acid active agent or non-omega-3 fattyacid nutritional supplement may be selected from the group consisting ofvitamin C, vitamin E, beta-carotene, magnesium, zinc (including as zincoxide), and copper (including as cupric oxide), and combinationsthereof, and at least one surface active agent, in an amount and acombination effective to cause said compositions to spontaneouslyself-micellize when in contact with an aqueous medium, thereby forming aplurality of stable micelles having a particle size within a range ofabout 1 to about 10 μm.

In certain embodiments, the compositions comprise at least one surfaceactive agent, at least one omega-3 fatty acid, and a non-omega-3 fattyacid active agent, wherein the omega-3 fatty acid (in either thetriglyceride, ester, or free fatty acid form) is eicosapentaenoic acid(EPA) (in either the triglyceride, ester, or free fatty acid form) andwherein the non-omega-3 fatty acid active agent is selected from thegroup consisting of vitamin C, vitamin E, beta-carotene, magnesium, zinc(including as zinc oxide), and copper (including as cupric oxide), andcombinations thereof.

In certain embodiments, the compositions comprise at least one surfaceactive agent, a first omega-3 fatty acid, a second omega-3 fatty acid,and a non-omega-3 fatty acid active agent, wherein the first omega-3fatty acid (in either the triglyceride, ester, or free fatty acid form)is EPA and the second omega-3 fatty acid (in either the triglyceride,ester, or free fatty acid form) is DHA, wherein the ratio of the amountof EPA:DHA is 1:X, where 0<X<1 and wherein the non-omega-3 fatty acidactive agent is selected from the group consisting of vitamin C, vitaminE, beta-carotene, magnesium, zinc (including as zinc oxide), and copper(including as cupric oxide), arachidonic acid (ARA), linoleic acid (LA),gamma-linolenic acid (GLA) or the omega-6 isomer of DPA and combinationsthereof.

Abnormal endothelial cell migration and proliferation can lead toadherent neovascularization. In the wet form of age-related maculardegeneration (AMD), neovascularization can create severe visionproblems. Current treatments include the use of known anti-angiogenesismedications. While not wishing to be bound to any particular theories,it is believed that compositions comprising omega-3 DPA can preventangiogenesis through the vascular endothelial growth factor (VEGF)pathway.

In at least one embodiment, a method is provided for treating AMD byadministering, to a patient in need thereof, a therapeutically effectiveamount of a composition comprising omega-3 DPA fatty acid (either in thetriglyceride, ester or free fatty acid ester form) and at least onesurface active agent, in an amount and a combination effective to causesaid compositions to spontaneously self-micellize when in contact withan aqueous medium, thereby forming a plurality of stable micelles havinga particle size within a range of about 1 μm to about 10 μm.

In at least one embodiment, a method is provided for treating fatmalabsorption syndromes by administering the compositions describedherein to a human in need of such administration. The human can be anadult, an adolescent, or and infant, such as for example a neonate.

In at least one embodiment, a method is provided for treating fatmalabsorption syndrome by administering the compositions describedherein to a human suffering from SBS or Barrett's Syndrome.

In at least one embodiment, a method is provided for treating sicklecell disease by administering the compositions described herein to ahuman in need of such administration. The human can be an adult, achild, an adolescent, or and infant, such as for example a neonate.

In at least one embodiment, a method is provided for treating NAFLDand/or NASH by administering the compositions described herein to ahuman in need of such administration. The human can be an adult, achild, an adolescent, or and infant, such as for example a neonate.

In at least one embodiment, a method is provided for treatingneurodegenerative diseases, such as for example, Parkinson's Disease(PD), Alzheimer's Disease (AD), Multiple Sclerosis (MS), Epilepsy, andAmyotrophic Lateral Sclerosis (ALS), by administering the compositionsdescribed herein to a human in need of such administration. The humancan be an adult, a child, an adolescent, or and infant, such as forexample a neonate.

In at least one embodiment of the invention, a method is provided forpreventing or treating Alzheimer's disease by administering thecompositions described herein to a subject in need of suchadministration, wherein said compositions may include at least onefat-soluble vitamin, at least one omega-3 fatty acid (in triglyceride,ester, or free fatty acid form), one or more minerals, such asmagnesium, manganese, zinc, copper, selenium, and combinations thereof,and at least one surface active agent in an amount and a combinationeffective to cause said compositions to spontaneously self-micellizewhen in contact with an aqueous medium, thereby forming a plurality ofstable micelles having a particle size within a range of about 1 μm toabout 10 μm.

In at least one embodiment of the invention, a method is provided forpreventing or treating age-related macular degeneration by administeringthe compositions described herein to a subject in need of suchadministration, wherein said compositions may include at least onefat-soluble vitamin, an omega-3 fatty acid (in triglyceride, ester, orfree fatty acid form, and one or more minerals, such as magnesium,manganese, zinc, copper, selenium, and combinations thereof, and atleast one surface active agent in an amount and a combination effectiveto cause said compositions to spontaneously self-micellize when incontact with an aqueous medium, thereby forming a plurality of stablemicelles having a particle size within a range of about 1 μm to about 10μm.

In at least one embodiment of the invention, a method is provided forpreventing or treating diseases by administering the compositionsdescribed herein to a subject in need of such administration, whereinsaid compositions may include at least one fat-soluble vitamin, anomega-3 fatty acid (in triglyceride, ester, or free fatty acid form),and one or more minerals, such as magnesium, manganese, zinc, copper,selenium, and combinations thereof, and at least one surface activeagent in an amount and a combination effective to cause saidcompositions to spontaneously self-micellize when in contact with anaqueous medium, thereby forming a plurality of stable micelles having aparticle size within a range of about 1 μm to about 10 μm., and whereinsaid diseases may include one or more of metabolic syndrome, chronicinflammation, hypovitaminosis D, zinc deficiency, Crohn's disease,diseases related to calcium deficiency, Alzheimer's disease, age-relatedmacular degeneration, short bowel syndrome, sickle cell disease,irritable bowel syndrome, clotting disorders, hypertriglyceridemia, PSC,non-alcoholic fatty liver disease, epilepsy, age-related dementia,diabetic retinopathy, insulin resistance, diabetes, muscular dystrophy,microvillus inclusion disease (also known as Davidson's disease,congenital microvillus atrophy), intestinal epithelial dysplasia (IED),also known as tufting enteropathy, syndromic diarrhea (SD), also knownas phenotypic diarrhea (PD) or tricho-hepato-enteric syndrome (THE), andcystic fibrosis.

In at least one embodiment of the invention, a method is provided fortreating short bowel syndrome by administering the compositionsdescribed herein to a subject in need of such administration, whereinsaid compositions may include at least one fat-soluble vitamin, anomega-3 fatty acid (in triglyceride, ester, or free fatty acid form),and one or more minerals, such as magnesium, manganese, zinc, copper,selenium, and combinations thereof, and an omega-6 fatty acid (intriglyceride, ester, or free fatty acid form), for example arachidonicacid (ARA), linoleic acid (LA), gamma-linolenic acid (GLA) or theomega-6 isomer of DPA, and at least one surface active agent in anamount and a combination effective to cause said compositions tospontaneously self-micellize when in contact with an aqueous medium,thereby forming a plurality of stable micelles having a particle sizewithin a range of about 1 μm to about 10 μm.

In certain embodiments, the compositions described herein self-micellizein an aqueous medium. In certain other embodiments, the aqueous mediumis water. In certain other embodiments, the aqueous medium has an acidicpH. In certain other embodiments, the aqueous medium is 0.1N HCl.

In certain embodiments, the compositions described herein self-micellizein an aqueous medium, wherein the micelles have a diameter from about 1μm to about 10 μm. In certain embodiments, the compositions describedherein self-micellizes in an aqueous medium having an acidic pH, whereinthe micelles have a diameter from about 1 μm to about 10 μm. In certainother embodiments, the compositions described herein self-micellizes in0.1N HCL, wherein the micelles have a diameter from about 1 μm to about10 μm. In certain embodiments, the micelles may have an average diameterof about 1, 2, 3, 4, 5, 6, 7, 8, 9, or 10 μm.

In certain embodiments, the compositions described herein minimize oreliminate at least one side effect when compared to the administrationof a composition comprising omega-3 fatty acids known to one of skill inthe art. Non-limiting examples of the side effects includeregurgitation, frequency of burping, gastroesophageal reflux disease(GERD), bloating, increased intestinal gas, fish taste, fishy breath,fish smell, nausea, diarrhea, or combinations thereof.

In certain embodiments, the compositions described herein comprised-limonene or natural orange oil. Such compositions can minimize oreliminate at least one side effect from the administration of acomposition of the present disclosure when compared to theadministration of a composition comprising omega-3 fatty acidssubstantially free of d-limonene or natural orange oil. Non-limitingexamples of the side effects include regurgitation, frequency ofburping, gastroesophageal reflux disease (GERD), bloating, increasedintestinal gas, fish taste, fishy breath, fish smell, nausea, diarrhea,or combinations thereof.

Certain embodiments of the compositions described herein can beadministered to a human subject in need of such administration inconjunction with a non-omega-3 fatty acid nutritional supplement or anon-omega-3 fatty acid active agent for the treatment of maculardegeneration. Non-limiting examples of non-omega-3 fatty acidnutritional supplements may include a combination of vitamin C, vitaminE, beta-carotene, zinc, copper, magnesium, manganese, calcium, vitaminA, vitamin D, vitamin K (includes K1 and/or K2), lutein and zeaxanthin.

Certain embodiments of the compositions described herein can beadministered to a human subject in need of such administration inconjunction with a non-omega-3 fatty acid active agent for the treatmentof SBS. Non-limiting examples of such non-omega-3 fatty acid activeagents can include L-Glutamine (NUTRESTORE®), recombinant somatotropin(ZORBTIVE®), and teduglutide (GATTEX®). Other such non-omega-3 fattyacid active agents may include conjugated bile acids or opium tincture.

Certain embodiments of the compositions described herein can beadministered to a human subject in need of such administration, eitheralone, or in conjunction with a non-omega-3 fatty acid nutritionalsupplement or a non-omega-3 fatty acid active agent. For example, (1) inthe treatment of a fat malabsorption syndrome, e.g. short bowelsyndrome, a composition within the purview of the present invention willinclude substantially pure DHA, or a combination of substantially pureDHA and arachidonic acid in combination with a surface active agentwhich is a combination including about 31% of Polysorbate 80 and about0.7% Poloxamer 237; (2) in the treatment of hypertriglyceridemia orhypocholesteremia, a composition within the purview of the presentinvention will include a combination of EPA and DHA, or a compositionincluding EPA and a therapeutically effective amount of one or morestatins, in combination with a surface active agent which is acombination including about 31% of Polysorbate 80 and about 0.7%Poloxamer 237; (3) in the treatment of non-alcoholic fatty liver disease(NAFLD) or non-alcoholic steatohepatitis (NASH), a composition withinthe purview of the present invention will include substantially pureDHA, in combination with a surface active agent which is a combinationincluding about 31% of Polysorbate 80 and about 0.7% Poloxamer 237; (4)in the treatment of sickle cell disease (SCD), a composition within thepurview of the present invention will include substantially pure DHAalone, substantially pure omega-3 DPA alone, or a combination of saidDHA and DPA, in combination with a surface active agent which is acombination including about 31% of Polysorbate 80 and about 0.7%Poloxamer 237, in a therapeutically effective amount to treat sicklecell disease symptoms, such as vaso-occlusive events, sequestrationcrisis, avascular necrosis, stroke, and sickle cell anemia, among otherknown maladies, by decreasing the aggregation of platelets, red bloodcells, and/or white blood cells.

Certain embodiments of the compositions described herein can beadministered to a human subject in need of such administration inconjunction with a non-omega-3 fatty acid nutritional supplement or anon-omega-3 fatty acid active agent for the treatment of sickle celldisease. Non-limiting examples of non-omega-3 fatty acid active agentscan include pain medications, antibiotics, a therapeutically effectiveamount of a non-steroidal anti-inflammatory medication, illustrated by,albeit not limited to a particular dosage of acetysalicylic acid(aspirin), hydroxyurea, and anti-inflammatory medications. Non-limitingexamples of non-omega-3 fatty acid nutritional supplements may include acombination of folic acid, vitamin C, vitamin E, beta-carotene, zinc,copper, magnesium, manganese, calcium, vitamin A, vitamin D, vitamin K(includes K1 and/or K2), lutein and zeaxanthin.

Certain embodiments of the compositions described herein can beadministered to a human subject in need of such administration inconjunction with a non-omega-3 fatty acid nutritional supplement or anon-omega-3 fatty acid active agent for the treatment of NAFLD and/orNASH. Non-limiting examples of non-omega-3 fatty acid active agents caninclude, lipid lowering or cholesterol lowering agents selected from thegroup consisting of cholesterol absorption inhibitors, bile acidsequestrants/resins, statins, niacin and derivatives, MTP inhibitors,fibrates and CETP inhibitors, insulin sensitizers, hypolipidemics,anti-inflammatory medications, and thiazolidinediones.

Certain embodiments of the compositions described herein can beadministered to a human subject in need of such administration inconjunction with a non-omega-3 fatty acid nutritional supplement or anon-omega-3 fatty acid active agent for the treatment of PSC or AlagilleSyndrome. Non-limiting examples of non-omega-3 fatty acid active agentscan, upon approval by a respective regulatory agency, may include6-alpha-ethylchenodeoxycholic acid and salts thereof, and/or(4R,5R)-1-[[4-[[4-[3,3-dibutyl-7-(dimethylamino)-2,3,4,5-tetrahydro-4-hydroxy-1,1-dioxido-1-benzothiepin-5-yl]phenoxy]methyl]phenyl]methyl]-4-aza-1-azoniabicyclo[2.2.2]octanechloride.

Certain embodiments of the compositions described herein can beadministered to a human subject in need of such administration inconjunction with a non-omega-3 fatty acid nutritional supplement or anon-omega-3 fatty acid active agent for the treatment of aneurodegenerative disease. Non-limiting examples of non-omega-3 fattyacid active agents can include medications generally approved by ahealth regulatory body and prescribed by physicians for a particularneurodegenerative disease, such as for example, PD, AD, MS, Epilepsy,and ALS.

In certain embodiments, compositions including substantially pure DHA incombination with at least one surface active agent, as described herein,effective to spontaneously form micelles, or alternatively, pre-formedmicelles produced therefrom, are effective for the treatment ofneurodevelopmental psychiatric disorders, such as Attention DeficitDisorder (ADD) and Attention Deficit Hyperactivity Disorder (ADHD).

In at least one embodiment, the compositions described herein can bemanufactured in an oil-filled inert, non-reactive vessel, non-limitingexamples of which include glass vials and Teflon® coated vials; with orwithout light penetrating inhibitors, including for example ambercolored vials. The composition in said oil-filled vial can beadministered to an adult human, adolescent, or infant, in particular toa neonate, in need of such administration, by adding an appropriateamount of saline or other pharmaceutically acceptable solutions to saidvial, thereby initiating the self-micellization process which results inthe formation of the pre-formed micelles as defined herein. Thehomogenous solution, containing the pre-formed micelles, can then beeither administered intravenously or added to infant formula for oralingestion When administered orally, the compositions described hereinmay be administered in the form of a gel or liquid filled capsule.

Also provided are kits comprising compositions described herein as oneor more unit dosage forms together with instructions on using the dosageforms. In certain embodiments, the dosage forms described herein can bepackaged as unit doses, such as for example, glass crimp or snap topvials with instructions for using the unit dosage forms. For example,the instructions can be provided as a package insert or directly on alabel attached to the vial, which can be made of clear or amber glass.The instructions can include, for example, dosing frequencyrecommendations, administration of the dosage forms with or withoutfood, and the active ingredients comprising the dosage forms.

In certain embodiments kits are provided, wherein certain dosage formscomprising the compositions described herein can be packaged togetherwith other non-omega-3 fatty acid nutritional supplements or activeagents for the treatment of AMD. The kit(s) comprise one or more unitdosage forms of certain embodiments of the compositions described hereintogether with one or more unit dosage forms comprising the non-omega-3fatty acid nutritional supplement or active agent for the treatment ofAMD together with instructions on using the dosage forms. Non-limitingexamples of non-omega-3 fatty acid nutritional supplements include, forexample, a combination of vitamin C, vitamin E, beta-carotene, zinc,copper, magnesium, manganese, calcium, vitamin A, vitamin D, vitamins K1and/or K2, lutein and zeaxanthin. Non-limiting examples of non-omega-3fatty acid active agents include, for example, verteporfin; theantioxidant carotenoids crocin and crocetin, as found in, for example,Saffron (Crocus sativus); and inhibitors of angiogenesis, such as forexample, BEVACIZUMAB (AVASTIN®), RANIBIZUMAB (LUCENTIS®), PEGAPTANIB(MACUGEN®) AND AFLIBERCEPT (EYLEA®).

Certain embodiments provide for a functional food(s) for treating AMDcomprising the compositions described herein.

Certain embodiments provide methods of treating AMD by administering afunctional food comprising the compositions described herein.

The instructions can include, for example, dosing frequency,administration of the dosage forms with or without food, the activeingredients comprising the dosage forms, and the fat malabsorptionsyndromes that would benefit from administration of the dosage forms.

In certain embodiments kits are provided, wherein certain dosage formscomprising the compositions described herein can be packaged togetherwith other non-omega-3 fatty acid nutritional supplements for thetreatment of a fat malabsorption syndrome. The kit(s) comprise one ormore unit dosage forms of certain embodiments of the compositionsdescribed herein together with one or more unit dosage forms comprisingthe non-omega-3 fatty acid nutritional supplements for the treatment ofa fat malabsorption syndrome together with instructions on using thedosage forms.

Certain embodiments provide for a functional food(s) for treating a fatmalabsorption syndrome comprising the compositions described herein.

Certain embodiments provide methods of treating a fat malabsorptionsyndrome by administering a functional food comprising the compositionsdescribed herein.

In certain embodiments kits are provided, wherein certain dosage formscomprising the compositions described herein can be packaged togetherwith other non-omega-3 fatty acid nutritional supplements or activeagents for the treatment of sickle cell disease. The kit(s) comprise oneor more unit dosage forms of certain embodiments of the compositionsdescribed herein together with one or more unit dosage forms comprisingthe non-omega-3 fatty acid nutritional supplement or active agent forthe treatment of sickle cell disease together with instructions on usingthe dosage forms. Non-limiting examples of non-omega-3 fatty acid activeagents and nutritional supplements that can be provided in the kitsinclude pain medications, antibiotics, folic acid, hydroxyurea, andanti-inflammatory medications

Certain embodiments provide for a functional food(s) for treating sicklecell disease comprising the compositions described herein.

Certain embodiments provide methods of treating sickle cell disease byadministering a functional food comprising the compositions describedherein.

In certain embodiments kits are provided, wherein certain dosage formscomprising the compositions described herein can be packaged togetherwith other non-omega-3 fatty acid nutritional supplements or activeagents for the treatment of NAFLD and/or NASH. The kit(s) comprise oneor more unit dosage forms of certain embodiments of the compositionsdescribed herein together with one or more unit dosage forms comprisingthe non-omega-3 fatty acid nutritional supplement or active agent forthe treatment of NAFLD and/or NASH together with instructions on usingthe dosage forms. Non-limiting examples of non-omega-3 fatty acid activeagents that can be provided in the kits include, lipid lowering orcholesterol lowering agents selected from the group consisting ofcholesterol absorption inhibitors, bile acid sequestrants/resins,statins, niacin and derivatives, MTP inhibitors, fibrates and CETPinhibitors, insulin sensitizers, hypolipidemics, anti-inflammatorymedications, and thiazolidinediones.

Certain embodiments provide for a functional food(s) for treating NAFLDand/or NASH comprising the compositions described herein. Certainembodiments provide methods of treating NAFLD and/or NASH byadministering a functional food comprising the compositions describedherein.

In certain embodiments kits are provided, wherein certain dosage formscomprising the compositions described herein can be packaged togetherwith other non-omega-3 fatty acid nutritional supplements or activeagents for the treatment of PSC. The kit(s) comprise one or more unitdosage forms of certain embodiments of the compositions described hereintogether with one or more unit dosage forms comprising the non-omega-3fatty acid nutritional supplement or active agent for the treatment ofPSC together with instructions on using the dosage forms. Non-limitingexamples of non-omega-3 fatty acid active agents that can, upon approvalby a respective regulatory agency, be provided in the kits include6-alpha-ethylchenodeoxycholic and/or(4R,5R)-1-[[4-[[4-[3,3-Dibutyl-7-(dimethylamino)-2,3,4,5-tetrahydro-4-hydroxy-1,1-dioxido-1-benzothiepin-5-yl]phenoxy]methyl]phenyl]methyl]-4-aza-1-azoniabicyclo[2.2.2]octaneChloride.

Certain embodiments provide for a functional food(s) for treating PSCcomprising the compositions described herein.

Certain embodiments provide methods of treating PSC by administering afunctional food comprising the compositions described herein.

In certain embodiments kits are provided, wherein certain dosage formscomprising the compositions described herein can be packaged togetherwith other non-omega-3 fatty acid nutritional supplements or activeagents for the treatment of neurodegenerative disease. The kit(s)comprise one or more unit dosage forms of certain embodiments of thecompositions described herein together with one or more unit dosageforms comprising the non-omega-3 fatty acid nutritional supplement oractive agent for the treatment of a neurodegenerative disease togetherwith instructions on using the dosage forms. Non-limiting examples ofnon-omega-3 fatty acid active agents that can be provided in the kitsinclude medications generally approved by a health regulatory body andprescribed by physicians for a particular neurodegenerative disease.

Certain embodiments provide for a functional food(s) for treatingneurodegenerative diseases, such as for example, PD, AD, MS, Epilepsy,or ALS, comprising the compositions described herein.

Certain embodiments provide methods of treating neurodegenerativediseases, such as for example, PD, AD, MS, Epilepsy, or ALS, byadministering a functional food comprising the compositions describedherein.

BRIEF DESCRIPTION OF THE DRAWINGS

FIGS. 1A-1D illustrate plasma fatty acid profiles (mol %) in the SBStreatment group (dark gray) versus Control (light gray) groups. DHA:Docosahexaenoic acid; EPA: Eicosapentaenoic acid; AA: Arachidonic acid,LA, Linoleic acid. Sample time 0=day of surgery; 1=postoperative day(POD) 1/AM; 2=POD 1/PM; 3=POD 2/AM; 4=POD 2/PM; 5=POD 3/AM; 6=POD 3/PM;7=POD 4/AM.

FIGS. 2A-2D illustrate intestinal morphology before and after surgicalinduction of SBS and DHA treatment. SBS: Short bowel syndrome; DHA:Docosahexaenoic acid; Black bars represent tissue morphometry fromsamples obtained at the time of surgery (Day 0); Striped bars representtissue morphometry from samples obtained at the end of the studyprotocol (Final).

FIG. 3 shows the mean body weight on the day of sacrifice of the micebetween each of the Control, Second Treatment Group and Third TreatmentGroup.

FIG. 4 shows α-SMA mRNA gene expression in mice in each of the control,NAFLD and NASH groups treated with either Vehicle or Composition 1 atthe end of the treatment period.

FIG. 5 shows plasma alanine transaminase (ALT) levels in mice betweeneach of the control, NAFLD and NASH group treated with either Vehicle orComposition at the end of the treatment period.

FIG. 6 shows the mean liver weight on the day of sacrifice of the micebetween each of the treatment groups and the control.

FIG. 7 shows the mean liver to body weight ratio on the day of sacrificeof the mice between each of the treatment groups and the control.

FIG. 8 shows whole blood glucose in mice between each of the control,NAFLD and NASH group treated with either Vehicle or Composition at theend of the treatment period.

FIG. 9 shows liver triglyceride levels in mice between each of thecontrol, NAFLD and NASH.

FIGS. 10 A and 10B show the mean individual EPA (A) and DHA (B) totallipid concentration-time profiles (baseline-adjusted change) after asingle dose of SC401 during fed and fasting conditions.

FIGS. 11A and 11B show the mean individual EPA (A) and DHA (B) freefatty acid concentration-time profiles (baseline-adjusted change) aftera single dose of SC401 during fed and fasting conditions.

FIG. 12 shows mean EPA and DHA total lipid plasma concentration profiles(μg/ml) (baseline-adjusted) after administration of a single dose (doseadjusted) of SC401 and Lovaza® in fasted conditions.

DETAILED DESCRIPTION

While the present invention is capable of being embodied in variousforms, the description below of several embodiments is made with theunderstanding that the present disclosure is to be considered as anexemplification of the invention, and is not intended to limit theinvention to the specific embodiments illustrated. Headings are providedfor convenience only and are not to be construed to limit the inventionin any manner. Embodiments illustrated under any heading may be combinedwith embodiments illustrated under any other heading.

The various embodiments of the invention described herein may suitablycomprise, consist essentially of, or consist of, at least one surfaceactive agent, at least one omega-3 fatty acid (in either thetriglyceride, ester, or free fatty acid form).

Certain embodiments of the invention may suitably comprise, consistessentially of, or consist of at least one surface active agent and afirst and second omega-3 fatty acid, wherein the first omega-3 fattyacid (in either the triglyceride, ester, or free fatty acid form) isselected from the group consisting of hexadecatrienoic acid, α-linolenicacid, stearidonic acid, eicosatrienoic acid, eicosapentaenoic acid,heneicosapentaenoic acid, docosapentaenoic acid, docosahexaenoic acid,tetracosapentaenoic acid, tetracosahexaenoic acid, and combinationsthereof and the second omega-3 fatty acid (in either the triglyceride,ester, or free fatty acid form) is selected from the group consisting ofhexadecatrienoic acid, α-linolenic acid, stearidonic acid,eicosatrienoic acid, eicosapentaenoic acid, heneicosapentaenoic acid,docosapentaenoic acid, docosahexaenoic acid, tetracosapentaenoic acid,tetracosahexaenoic acid, and combinations thereof, wherein the first andsecond omega-3 fatty acids (in either the triglyceride, ester, or freefatty acid form) are different and wherein the ratio of the firstomega-3 fatty acid (in either the triglyceride, ester, or free fattyacid form):the second omega-3 fatty acid (in either the triglyceride,ester, or free fatty acid form) is 1:X, where 0<X<0.3 or 0.5<X<1.

Certain embodiments of the invention may suitably comprise, consistessentially of, or consist of at least one surface active agent and afirst and second omega-3 fatty acid, wherein the first omega-3 fattyacid (in either the triglyceride, ester, or free fatty acid form) is DHAand the second omega-3 fatty acid (in either the triglyceride, ester, orfree fatty acid form) is selected from the group consisting ofhexadecatrienoic acid, α-linolenic acid, stearidonic acid,eicosatrienoic acid, eicosapentaenoic acid, heneicosapentaenoic acid,docosapentaenoic acid, tetracosapentaenoic acid, tetracosahexaenoicacid, and combinations thereof, wherein the ratio of the amount of DHAto the second omega-3 fatty acid is 1:X, where 0<X<0.3 or 0.5<X<1.

Certain embodiments of the invention may suitably comprise, consistessentially of, or consist of at least one surface active agent and afirst and second omega-3 fatty acid, wherein the first omega-3 fattyacid (in either the triglyceride, ester, or free fatty acid form) is EPAand the second omega-3 fatty acid (in either the triglyceride, ester, orfree fatty acid form) is selected from the group consisting ofhexadecatrienoic acid, α-linolenic acid, stearidonic acid,eicosatrienoic acid, heneicosapentaenoic acid, docosapentaenoic acid,docosahexaenoic acid, tetracosapentaenoic acid, tetracosahexaenoic acid,and combinations thereof, wherein the ratio of the amount of EPA to thesecond omega-3 fatty acid is 1:X, where 0<X<0.3 or 0.5<X<1.

Definitions

As used herein, the term “composition(s)” or “formulation(s)” includestherapeutic and dietary compositions including, but not limited to, adietary supplement, nutraceutical formulation, or pharmaceuticalformulation. Further, the terms composition, dietary supplement,nutraceutical formulation, and pharmaceutical formulation are usedinterchangeably herein.

As used herein, the term “EPA” refers inclusively to(5Z,8Z,11Z,14Z,17Z)-eicosa-5,8,11,14,17-pentaenoic acid or derivativesthereof, including alkyl esters, such as, for example, the ethyl ester.

As used herein, the term “DHA” inclusively refers to(4Z,7Z,10Z,13Z,16Z,19Z)-docosa-4,7,10,13,16,19-hexaenoic acid orderivatives thereof, including alkyl esters, such as, for example, theethyl ester.

As used herein, the term “DPA” refers inclusively to docosapentaenoicacid (DPA), wherein “omega-3 DPA” refers to the omega-3 isomerall-cis-7,10,13,16,19-docosapentaenoic acid (clupanodonic acid) and“omega-6 DPA” refers to the omega-6 isomerall-cis-4,7,10,13,16-docosapentaenoic acid (osbond acid).

As used herein, the term “ARA” refers inclusively to(5Z,8Z,11Z,14-Z)-Icosa-5,8,11,14-tetraenoic acid or derivatives thereof,including alkyl esters, such as, for example, the ethyl ester.

As used herein, the term linoleic acid “LA”, refers inclusively to(9Z,12Z)-9,12-Octadecadienoic acid or derivatives thereof, includingalkyl esters, such as, for example, the ethyl ester.

As used herein, the term “GLA” refers inclusively toall-cis-6,9,12-octadecatrienoic acid or derivatives thereof, includingalkyl esters, such as, for example, the ethyl ester.

As used herein, the term “micelle” (plural micelles, micella, pluralityof micelles, or micellae) refers to an aggregate of molecules, that haveassembled into a substantially spherical core/shell architecture, andare suspended in an aqueous phase. A typical micelle in aqueous solutionforms an aggregate with the hydrophilic “head” regions in contact withsurrounding solvent and/or in contact with the polar region of one ormore surface active agent(s), sequestering the hydrophobic regions inthe micelle center. Micelles are approximately spherical in shape.

The term “self-micellizes” or “self-micellization” as used herein refersto the process in which micelles are formed in an aqueous medium withoutthe introduction of energy, including agitation or shearing.

As used herein, the term “aqueous medium” refers to any solution orsuspension, that comprises in part or in whole water, including forexample, without limitation, water by itself; phosphate buffered salineat about pH 7.4, soft-drinks, illustrated by, albeit not limited toSprite®, apple juice, G-2® fruit punch, infant formula or any equivalentmammalian mother's milk substitutes or analogues, intravenous fluid, andmilk and milk products, including chocolate milk. In certainembodiments, an aqueous medium comprises at least one fluid having atleast one fatty acid, carbohydrate, lipid, sugar, or combinationsthereof. In certain embodiments, an aqueous medium comprises at leastone fluid having an acidic pH. In certain other embodiments, an aqueousmedium comprises a biological fluid such as, for example and withoutlimitation, stomach acid. In other embodiments, the aqueous mediumcomprises simulated stomach acid comprising 0.1N HCl.

As used herein, the term “free fatty acid” refers to one or morepolyunsaturated fatty acids that have not been modified or do not haveany other groups attached.

As used herein, the term “ester” refers to the replacement of thehydrogen in the carboxylic acid group of a polyunsaturated fatty acidmolecule with another substituent. Typical esters are known to those inthe art, a discussion of which is provided by Higuchi, T. et al.,Pro-drugs as Novel Delivery Systems, Vol. 14, A.C.S. Symposium Series,Bioreversible Carriers in Drug Design, Ed. Edward B. Roche, Amer.Pharma. Assoc., Pergamon Press (1987), and Protective Groups in OrganicChemistry, McOmie ed., Plenum Press, New York (1973), each of which isincorporated herein by reference in their entirety. Examples of commonesters include methyl, ethyl, trichloroethyl, propyl, butyl, pentyl,tert-butyl, benzyl, nitrobenzyl, methoxybenzyl, benzhydryl,monoglyceride, diglyceride, triglyceride.

As used herein, the term “monoglyceride” refers to a fatty acid chain,such as DHA or EPA molecule, covalently bonded to a glycerol moleculethrough an ester linkage. As used herein, the term “diglyceride” refersto a fatty acid chain such as DHA or EPA, covalently bonded to aglycerol molecule through an ester linkage, wherein the glycerolmolecule is further bonded to one additional fatty acid chain, which mayor may not be DHA or EPA, through one additional ester linkage. As usedherein, the term “triglyceride” refers to a fatty acid chain, such asDHA or EPA, covalently bonded to a glycerol molecule through an esterlinkage, wherein the glycerol molecule is further bonded to twoadditional fatty acid chains, either or both of which may or may not beDHA or EPA, through two additional ester linkages.

As used herein, the term “terpene” refers to the large and diverse classof organic compounds produced by a variety of plants, particularlyconifers. When terpenes are modified chemically, such as by oxidation orrearrangement of the carbon skeleton, the resulting compounds aregenerally referred to as “terpenoids” (e.g., carvone). Terpenes andterpenoids are the primary constituents of the essential oils of manytypes of plants and flowers.

As used herein, the terms “tocopherol”, and “tocotrienol”, and “vitaminE” refer to a set of fat-soluble vitamins with antioxidant properties.For example, the term “tocotrienol” refers to the family of tocotrienols“alpha-tocotrienol”, “beta-tocotrienol”, “gamma-tocotrienol” and“delta-tocotrienol”. Similarly, the term “tocopherol” refers to thefamily of tocopherols “alpha-tocopherol”, “beta-tocopherol”,“gamma-tocopherol” and “delta-tocopherol”.

As used herein, the term “antioxidant” refers to a molecule capable ofinhibiting the oxidation of other molecules. Oxidation is a chemicalreaction that transfers electrons or hydrogen from a substance to anoxidizing agent. Oxidation reactions can produce free radicals. In turn,these radicals can start chain reactions. When the chain reaction occursin a cell, it can cause damage or death to the cell. Antioxidantsterminate these chain reactions by removing free radical intermediates,and inhibit other oxidation reactions. They do this by being oxidizedthemselves, so antioxidants are often reducing agents such as thiols,ascorbic acid, or polyphenols. Exemplary antioxidants include rosemaryoil, ascorbic acid (vitamin C), glutathione, lipoic acid, uric acid,carotenes, melatonin, ubiquinol (coenzyme Q), α-tocopherol (vitamin E),acorbyl palmitate, butylated hydroxyanisole, butylated hydroxytoluene,monothioglycerol, and potassium metabisulfite.

As used herein, a “pharmaceutically acceptable carrier” refers to anynon-toxic amount of any substance suitable as a vehicle for delivering amolecule or composition to a suitable in vivo site of absorption.Examples of such carriers include, but are not limited to water,phosphate buffered saline (PBS), Ringer's solution, dextrose solution,serum-containing solutions, Hank's solution and other aqueousphysiologically-balanced solutions.

As used herein, a “pharmaceutically acceptable preservative” includesbut is not limited to a non-toxic amount of potassium sorbate,methylparaben, propylparaben, benzoic acid and its salts, other estersof parahydroxybenzoic acid such as butylparaben, alcohols such as ethylor benzyl alcohol, phenolic compounds such as phenol, or quarternarycompounds such as benzalkonium chloride.

As used herein, a “coloring agent” provides coloration to thecomposition or dosage form. Such coloring agents include, for example,food grade dyes.

As used herein, the term “subject” refers to a mammal, including but notlimited to a dog, cat, horse, cow, pig, sheep, goat, chicken, rodent,primate or human. Subjects include animals such as house pets (e.g.,dogs, cats, and the like), agricultural stock subjects (e.g., cows,horses, pigs, chickens, etc.), laboratory subjects (e.g., mice, rats,rabbits, etc.), but are not so limited. The human subject may be apediatric, such as for example neonate, adult, or a geriatric subject.The human subject may be of either gender.

As used herein, an “effective amount” or “therapeutically effectiveamount” of a composition as described in some embodiments herein can bea quantity sufficient to achieve a desired therapeutic and/orprophylactic effect, for example, an amount which results in theprevention of, or a decrease in the symptoms associated with, a diseasethat is being treated. The amount of composition administered to thesubject, particularly one in need of the composition, can depend on thetype and severity of the disease and on the characteristics of theindividual, such as general health, age, sex, body weight and toleranceto drugs. A person skilled in the art will be able to determineappropriate dosages depending on these and other factors. Typically, aneffective amount of the compositions described herein can be sufficientfor achieving a therapeutic or prophylactic effect.

In conjunction with the determination of a “therapeutically effectiveamount” of a particular modality, it is within the purview of thepresent invention to determine the presence of such a “therapeuticallyeffective amount” by determining a quantity of a composition asdescribed in some embodiments herein to modulate biomarkers related to aparticular disease state. Such biomarkers may form a basis for surrogateendpoints for clinical trials, provide a basis for elucidating a betterunderstanding of disease pathogenesis, and for quantifying theeffectiveness of a therapeutic modality to down-regulate an inflammatoryresponse, thereby quantifying disease progression.

Illustrative, albeit non-limiting biomarkers for several of the diseasestates effectively treated by the compositions of the present inventionmay include:

SCD

D-Dimer, to assess the coagulation rate, Total white blood cells andplatelets, markers of inflammatory state, Lactate dehydrogenase (LDH),marker of intravascular hemolysis, Dense red blood cell fraction, markerof red blood cell sickling, Endothelin-1 (ET-1), soluble-VCAM-1(sVCAM-1), soluble P-selectin (sP-selectin), markers of endothelialactivation, interleukins (IL2, IL3, IL6, IL8 and IL10), urinarycysteinyl leucotriene E4 (28), and serum levels of prostaglandin-E2, CA15-3, soluble CD40 ligand, HSP-70, ferritin, angiopoietin 1 and 2,stromal derived factor 1, tumor necrosis factor-α and tumor necrosisfactor receptor-1.

NAFLD/NASH

CK18 M30, marker of NAFLD severity, IL-6, inflammatory state, Urineisoprostane, Oxidative stress, Livers spectroscopy, Fatty liverassessment

PSC

Calprotectin, Alkaline phosphatase—an elevated ALP is the most commonbiochemical abnormality in PSC, which often prompts clinicians toestablish a diagnosis, reduction of ALP over time is associated with afavorable prognosis, Aspartate to alanine aminotransferase ratio:Aspartate aminotransferase (AST) and alanine aminotransferase (ALT) aremarkers of hepatocellular injury or necrosis, and elevation of AST orALT is present in case of liver damage, Bilirubin: Bilirubin isroutinely used in clinical practice as a measure of extrahepaticobstruction, Immunoglobulin G4: Measure of inflammation, inflammationresults in fibrosis and deposition of connective tissue. PSC patientswith elevated IgG4 have been shown to have a high prevalence ofcirrhosis, and a shorter time to liver transplant, suggesting a moresevere liver disease course.

It is contemplated to elucidate such biomarkers for any of the diseasestates treated herein, and to determine a therapeutically effectiveamount of a composition as it relates to its ability to modulate thepathogenesis of said disease state.

The terms “dose unit,” “unit dose,” and “dosage unit,” as used herein,refer to a portion of a composition that contains an effective amount ofan active suitable for a single administration to provide, or contributeto, a therapeutic effect. Such dosage units may be administered one to aplurality (i.e., 1 to about 10, 1 to about 8, 1 to about 6, 1 to about 4or 1 to about 2) of times per day, or as many times as needed to elicita therapeutic response.

The term “food effect,” as used herein, refers to a relative differencein AUC (area under the curve), C_(max) (maximum plasma concentration),and/or T_(max) (time to maximum concentration) of an active substance,when said substance or a composition thereof, such as a tablet, acapsule or a liquid, is administered orally to a subject concomitantlywith food or in a fed state as compared to the same values when the samecomposition is administered in a fasted state. The food effect, F, iscalculated as:

F=(Y _(fed) −Y _(fasted))/Y _(fasted)

wherein Y_(fed) and Y_(fasted) are the found values of AUC, C_(max), orT_(max) in the fed and fasted state, respectively. A food effect, F, isgenerally established when F>1.

In general, the term “AUC” or “area under the plasma concentration-timecurve” is related to the total amount of an active measurable in thesystemic circulation following administration of a single dose. The AUCis a mathematical and visual representation of the aggregate amount ofthe active in the systemic circulation over a given period of time.Changes in the AUC need not necessarily reflect changes in the totalamount of the active absorbed but can reflect modifications in thekinetics of distribution, metabolism and excretion. Accordingly, theterm AUC as used herein refers to the total amount of omega-3 fattyacids (in either the triglyceride, ester, or free fatty acid form)measurable in the systemic circulation following administration of asingle dose of any of the compositions described herein.

The term “T_(max)” or “time of peak concentration” refers to the periodof time required to achieve peak plasma concentration of an active afteradministration of a single dose. Accordingly, the term T_(max) as usedherein refers to the period of time required to achieve peak plasmaconcentration of omega-3 fatty acid (in either the triglyceride, ester,or free fatty acid form) after administration of a single dose of any ofthe compositions described herein.

The term “C_(max)” or “peak concentration” is the highest concentrationof an active achieved in the blood plasma. Accordingly, the term C_(max)as used herein refers to the maximum concentration of omega-3 fattyacids (in either the triglyceride, ester, or free fatty acid form) afteradministration of a single dose of any of the compositions describedherein.

The term “substantially independent of a food effect,” or “substantiallyfree of food effect” as used herein, refers to a significant eliminationof the effect of food upon the absorption (e.g., F is about 0),following oral administration, of any of the compositions describedherein. In other words, the bioavailability of the omega-3 fatty acids(in either the triglyceride, ester, or free fatty acid form), asmeasured by the logarithm-transformed AUC_(0-∞), is bioequivalent i.e.,the 90% confidence interval should be within the acceptance range of 80%to 125% regardless of whether the compositions described herein areadministered with or without food. In certain embodiments, thepharmacological effects of administration of compositions describedherein are substantially independent of a food effect.

The term “reduced food effect,” as used herein, refers to a substantialreduction in the effect of food upon the absorption, following oraladministration, of any of the compositions described. In certainembodiments, the compositions described herein have a reduced foodeffect.

The term “concomitantly with food” or “administration in the fed state,”as used herein, refers to administration from about 30 minutes before ameal to about 1 hour after a meal.

Various modes of treatment or prevention of medical conditions asdescribed herein are intended to mean “substantial” or “substantially”,which includes total but also less than total treatment or prevention,and wherein some biologically or medically relevant result is achieved.A subject, such as a human subject, in need of treatment refers to asubject in need of treatment of a defined disease state or in need ofpreventative treatment (i.e., prophylaxis) of such a disease state.

The use of numerical values in the various quantitative values specifiedthroughout this disclosure, unless expressly indicated otherwise, arestated as approximations as though the minimum and maximum values withinthe stated ranges were both preceded by the word “about” unlessexpressly stated otherwise. In this manner, equivalent variations from astated value can be used to achieve substantially the same results asthe stated value. Also, the disclosure of ranges is intended as acontinuous range including every value between the minimum and maximumvalues recited as well as any ranges that can be formed by such values.Also disclosed herein are any and all ratios (and ranges of any suchratios) that can be formed by dividing a recited numeric value into anyother recited numeric value. Accordingly, the skilled person willappreciate that many such ratios, ranges, and ranges of ratios can beunambiguously derived from the numerical values presented herein and inall instances such ratios, ranges, and ranges of ratios representvarious embodiments of the present invention.

The term “functional food” as used herein means any edible or drinkablefoods or dietary components (e.g., infant formula, juices, milk, yogurt,butter, margarine, baking products) that are fortified or enhanced withany of the compositions described herein. The functional food can be,e.g., solid, liquid, semisolid, or a combination thereof. The term“functional food” also encompasses edible and drinkable nutritionalsupplements.

The term “hydrophilic-lipophilic balance” or “HLB,” as used herein,refers to the relative affinity of a substance or composition foraqueous and oily phases. HLB values can be calculated based on methodsand equations known to those of ordinary skill in the art, such as thosedescribed in U.S. Pat. No. 5,585,192. Substances or compositionsgenerally have an average HLB of about 6 to about 20.Hydrophilic-lipophilic balance values can be determined in a variety ofthe formulas or experimental methods provided, for example, in U.S. Pat.No. 5,585,192.

The term “substantially pure” as used herein means at least 80% pure.

Pharmaceutical Compositions

In at least one embodiment, a composition is provided, wherein thecomposition comprises at least one omega-3 fatty acid and at least onesurface active agent. In certain embodiments, the at least one omega-3fatty acid is selected from the group consisting of hexadecatrienoicacid, α-linolenic acid, stearidonic acid, eicosatrienoic acid,eicosapentaenoic acid, heneicosapentaenoic acid, docosapentaenoic acid,docosahexaenoic acid, tetracosapentaenoic acid, and tetracosahexaenoicacid. Certain embodiments comprise more than one omega-3 fatty acid,which can be selected from the group consisting of hexadecatrienoicacid, α-linolenic acid, stearidonic acid, eicosatrienoic acid,eicosapentaenoic acid, heneicosapentaenoic acid, docosapentaenoic acid,docosahexaenoic acid, tetracosapentaenoic acid, and tetracosahexaenoicacid. The omega-3 fatty acid components comprising the composition arepresent at, for example, 60%, 65%, 70%, 75%, 80%, 85%, 90% or 95%(wt/wt) of the composition. The omega-3 fatty acids used herein aretypically in the form of an oil.

In at least one embodiment, a composition is provided, wherein thecomposition comprises at least one isomer of docosapentaenoic acid (DPA)and at least one surface active agent, wherein the omega-3 DPA isomer isall-cis-7,10,13,16,19-docosapentaenoic acid (clupanodonic acid) or theomega-6 DPA isomer is all-cis-4,7,10,13,16-docosapentaenoic acid (osbondacid).

In certain embodiments, the compositions described herein comprise DHAat, for example, from 50% to 99% (wt/wt) of the compositions. Forexample, in certain embodiments, the DHA is present at 50%, 55%, 60%,65%, 70%, 75%, 80%, 85%, 90%, 95%, or 99% (wt/wt) of the compositions.In certain other embodiments, the DHA is present at least, for example,90% (wt/wt) of the compositions described herein. In still otherembodiments, the DHA is present at, at least, for example, 95% (wt/wt)of the compositions described herein.

In certain embodiments, the compositions described herein comprise EPAat, for example, from 0% to 50% (wt/wt) of the compositions. Forexample, in certain embodiments, the EPA is present at <1% (wt/wt) ofsaid compositions. In certain other embodiments, the EPA can be presentat, for example, 2%, 4%, 6%, 8%, 10%, 12%, 14%, 16%, 18%, 20%, 22%, 24%,26%, 28%, 30%, 32%, 34%, 36%, 38%, 40%, 42%, 44%, 46%, 48% or 50%(wt/wt) of said compositions. In still other embodiments, the EPA can bepresent at <5% (wt/wt) of said compositions. For example, in certainembodiments, the EPA can be present at 0%, 0.1%, 0.2%, 0.3%, 0.4%, 0.5%,0.6%, 0.7%, 0.8%, 0.9%, 1%, 1.5%, 2%, 2.5%, 3%, 3.5%, 4%, or 4.5%(wt/wt) of said compositions.

In certain embodiments, the compositions described herein comprise anomega-3 fatty acid other than DHA and EPA at, for example, from >0% to10% (wt/wt) of said compositions. For example, said omega-3 fatty acidother than DHA and EPA can be present, in certain embodiments at 0.1%,0.2%, 0.3%, 0.4%, 0.5%, 0.6%, 0.7%, 0.8%, 0.9%, 1%, 1.5%, 2%, 2.5%, 3%,3.5%, 4%, 4.5%, 5%, 5.5%, 6.5%, 7%, 7.5%, 8%, 8.5%, 9%, 9.5% or 10%(wt/wt) of said compositions. In such embodiments, the omega-3 fattyacid other than EPA and DHA can be selected from the group consisting ofhexadecatrienoic acid, α-linolenic acid, stearidonic acid,eicosatrienoic acid, heneicosapentaenoic acid, docosapentaenoic acid,tetracosapentaenoic acid, and tetracosahexaenoic acid. In certain suchembodiments, the third fatty acid other than DHA and EPA may be anomega-6 fatty acid, for example arachidonic acid (ARA), linoleic acid(LA), gamma-linolenic acid (GLA) or the omega-6 isomer of DPA.

In certain embodiments, the compositions described herein comprise 90%,5%, and 5% (wt/wt) of said compositions DHA, ARA, and EPA respectively.

In certain embodiments, the compositions described herein comprise 95%,5%, and <1% (wt/wt) of said compositions DHA, ARA, and EPA respectively.

The omega-3 fatty acid oils used in the manufacture of the compositionsdescribed herein are substantially pure. In some embodiments, theomega-3 fatty acid oils are at least, for example, 90% pure; and theremainder of the omega-3 fatty acids in the oil can comprise a mixtureof other omega-3 fatty acids of which one omega-3 fatty acid maypredominate the mixture. For example, if a composition comprises DHAoil, the DHA oil can be at least 90% DHA; and the remainder can comprisea mixture of omega-3 fatty acids, of which EPA can be the predominantomega-3 fatty acid and can be present at 5% of said composition.Accordingly, in omega-3 fatty acid oils, the ratio of the amount of thesubstantially pure omega-3 fatty acid, the first omega-3 fatty acid, tothe predominant omega-3 fatty acid in the remaining fraction of the oil,the second omega-3 fatty acid, is 1:X, where 0<X<0.3 or 0.5<X<1. Thus,in certain embodiments, X can be, for example, 0.001, 0.0015, 0.002,0.0025, 0.0055, 0.006, 0.0065, 0.007, 0.0075, 0.008, 0.0085, 0.009,0.0095, 0.01, 0.015, 0.02, 0.025, 0.03, 0.035, 0.04, 0.045, 0.05, 0.055,0.06, 0.065, 0.07, 0.075, 0.08, 0.085, 0.09, 0.095, 0.1, 0.15, 0.2,0.25, 0.3, 0.55, 0.6, 0.65, 0.7, 0.75, or 0.8.

Thus, in at least one embodiment, the composition described hereincomprises at least one surface active agent and a first and secondomega-3 fatty acid, wherein the first omega-3 fatty acid is DHA and thesecond omega-3 fatty acid EPA, wherein the ratio of the amount ofDHA:EPA is 1:X, where 0<X<0.3 or 0.5<X<1. In certain other embodiments,X can be, for example, 0.001, 0.0015, 0.002, 0.0025, 0.0055, 0.006,0.0065, 0.007, 0.0075, 0.008, 0.0085, 0.009, 0.0095, 0.01, 0.015, 0.02,0.025, 0.03, 0.035, 0.04, 0.045, 0.05, 0.055, 0.06, 0.065, 0.07, 0.075,0.08, 0.085, 0.09, 0.095, 0.1, 0.15, 0.2, 0.25, 0.55, 0.6, 0.65, 0.7,0.75, or 0.8.

In other embodiments, the compositions described herein comprise atleast one surface active agent and a first and second omega-3 fattyacid, wherein the first omega-3 fatty acid is EPA and the second omega-3fatty acid is DHA, wherein the ratio of the amount of EPA:DHA is 1:X,where 0<X<0.3 or 0.5<X<1. In certain other embodiments, X can be, forexample, 0.001, 0.0015, 0.002, 0.0025, 0.0055, 0.006, 0.0065, 0.007,0.0075, 0.008, 0.0085, 0.009, 0.0095, 0.01, 0.015, 0.02, 0.025, 0.03,0.055, 0.06, 0.065, 0.07, 0.075, 0.08, 0.085, 0.09, 0.095, 0.1, 0.15,0.2, 0.25, 0.55, 0.6, 0.65, 0.7, 0.75, or 0.8.

In certain embodiments, the compositions described herein can optionallycomprise a third omega fatty acid (either in the triglyceride, ester, orfree fatty acid form), wherein the third omega fatty acid is differentfrom the first and second omega fatty acids. In such compositions, theratio of the amount of the first omega fatty acid to the second andthird omega fatty acids is 1:X, where X is the combined amount of thesecond and third omega fatty acids and X is 0<X<1. For example, if acomposition comprises DHA oil, the DHA oil can be at least, for example,90% DHA (the first omega fatty acid); and the remaining 10% can comprisea mixture of omega fatty acids, of which EPA can be the predominantomega fatty acid (second omega fatty acid) and can be present at 5% ofsaid composition. The third omega fatty acid is selected from the groupconsisting of hexadecatrienoic acid, α-linolenic acid, stearidonic acid,eicosatrienoic acid, heneicosapentaenoic acid, docosapentaenoic acid,tetracosapentaenoic acid, and tetracosahexaenoic acid. In certainembodiments the third omega acid may be an omega-6 fatty acid.Accordingly, in such embodiments, if the first omega fatty acid is DHAand the second omega fatty acid is EPA, The ratio of the amount ofDHA:(EPA+third omega fatty acid) is 1:X, where X is the amount of EPAand the amount of the third omega fatty acid, and where 0<X<1. Incertain embodiments, X can be, for example, 0.001, 0.0015, 0.002,0.0025, 0.0055, 0.006, 0.0065, 0.007, 0.0075, 0.008, 0.0085, 0.009,0.0095, 0.01, 0.015, 0.02, 0.025, 0.03, 0.035, 0.04, 0.045, 0.05, 0.055,0.06, 0.065, 0.07, 0.075, 0.08, 0.085, 0.09, 0.095, 0.1, 0.15, 0.2,0.25, 0.3, 0.35, 0.4, 0.45, 0.5, 0.55, 0.6, 0.65, 0.7, 0.75, 0.8, 0.85,0.9, or 0.95. In certain embodiments, the third fatty acid can bearachidonic acid (ARA) (either in the triglyceride, ester, or free fattyacid form). In certain embodiments, the ARA is in the ethyl ester form.

In some embodiments, the omega-3 or omega-6 fatty acid compositionsdescribed herein are in either the triglyceride, ester, or free fattyacid form. Typically, but not necessarily, the omega-3 or omega-6 fattyacid is in the ester form, particularly in the ethyl ester form.

In certain embodiments, the compositions comprise at least one surfaceactive agent and a first and second omega-3 fatty acid, wherein thefirst omega-3 fatty acid (in either the triglyceride, ester, or freefatty acid form) is omega-3 docosapentaenoic acid (omega-3 DPA) and thesecond omega-3 fatty acid (in either the triglyceride, ester, or freefatty acid form) is selected from the group consisting ofdocosahexaenoic acid (DHA), eicosapentaenoic acid (EPA),hexadecatrienoic acid, α-linolenic acid, stearidonic acid,eicosatrienoic acid, eicosapentaenoic acid, heneicosapentaenoic acid,docosapentaenoic acid, tetracosapentaenoic acid, and tetracosahexaenoicacid and combinations thereof.

In certain embodiments, the compositions comprise at least one surfaceactive agent, an omega-6 fatty acid, and an omega-3 fatty acid, whereinthe omega-6 fatty acid (in either the triglyceride, ester, or free fattyacid form) is for example arachidonic acid (ARA), linoleic acid (LA),gamma-linolenic acid (GLA) or the omega-6 isomer of DPA, and the secondomega-3 fatty acid (in either the triglyceride, ester, or free fattyacid form) is selected from the group consisting of omega-3docosapentaenoic acid (omega-3 DPA), docosahexaenoic acid (DHA),eicosapentaenoic acid (EPA), hexadecatrienoic acid, α-linolenic acid,stearidonic acid, eicosatrienoic acid, eicosapentaenoic acid,heneicosapentaenoic acid, docosapentaenoic acid, tetracosapentaenoicacid, and tetracosahexaenoic acid and combinations thereof.

In certain embodiments the omega-3 fatty acid (in either thetriglyceride, ester, or free fatty acid form) may be DHA, for examplesubstantially pure DHA, in combination with arachidonic acid in a ratioof DHA to arachidonic acid of 1:1 to 20:1. In certain embodiments,α-linolenic acid may be included for its anti-inflammatory properties.

In certain embodiments, the compositions described herein are free offree fatty acids.

In certain embodiments, the compositions described herein are free of asecond medicament.

In certain embodiments, the at least one surface active agent isselected from the group consisting of nonionic surface active agents,cationic surface active agents, anionic surface active agents,zwitterionic surface active agents, or combinations thereof.

In some embodiments, the compositions include one or more non-ionicsurface active agents. Non-ionic surface active agents generally have ahydrophobic group and a reactive hydrogen atom, for example aliphaticalcohols, acids, amides and alkyl phenols, with alkylene oxides,especially ethylene oxide either alone or in combination with propyleneoxide. Examples of nonionic surfactant compounds include, but are notlimited to, polyoxyethylene glycol sorbitan alkyl esters, blockcopolymers of polyethylene glycol and polypropylene glycol, ethyleneglycol fatty acid esters, poly(ethylene glycol) fatty acid esters,propylene glycol fatty acid esters, poly(propylene glycol) fatty acidesters, glycol fatty acid esters, trimethylolpropane fatty acid esters,pentaerythritol fatty acid esters, glucoside derivatives, glycerin alkylether fatty acid esters, trimethylolpropane oxyethylene alkyl ethers,fatty acid amides, alkylolamides, alkylamine oxides, lanolin and itsderivatives, castor oil derivatives, hardened castor oil derivatives,sterols and its derivatives, polyoxyethylene alkyl ethers,polyoxyethylene alkyl allyl ethers, polyoxyethylene alkylamine,polyoxyethylene fatty acid amides, polyoxyethylene alkylolamides,polyoxyethylene diethanolamine fatty acid esters, polyoxyethylenetrimethylolpropane fatty acid esters, polyoxyethylene alkyl ether fattyacid esters, polyoxyethylene polyoxypropylene glycols, polyoxyethylenepolyoxypropylene alkyl ethers, polyoxyethylene polyoxypropylenepolyhydric alcohol ethers, glycerin fatty acid esters, polyglycerinfatty acid esters, polyoxyethylene glycerin fatty acid esters, sorbitanfatty acid esters, polyoxyethylene sorbitan fatty acid esters, sucrosefatty acid esters, or combinations thereof.

In certain embodiments, the surface active agents comprisepolyoxyethylene glycol sorbitan alkyl esters, block copolymers ofpolyethylene glycol and polypropylene glycol, or combinations thereof.

Examples of polyoxyethylene glycol sorbitan alkyl esters are typicallythe polysorbates. Polysorbates are a class of oily liquids derived fromPEG-ylated sorbitan (a derivative of sorbitol) esterified with fattyacids. Common brand names for polysorbates include Tween®. Tween®-20,Tween®-40, Tween®-60 and Tween®-80, for example, are available fromAkzoNobel (Strawinskylaan 2555 1077 ZZ, Amsterdam, the Netherlands).Exemplary polysorbates include polysorbate 20 (polyoxyethylene (20)sorbitan monolaurate), polysorbate 40 (polyoxyethylene (20) sorbitanmonopalmitate), polysorbate 60 (polyoxyethylene (20) sorbitanmonostearate), and polysorbate 80 (polyoxyethylene (20) sorbitanmonooleate).

Examples of block copolymers of polyethylene glycol and polypropyleneglycol include the poloxamers. Poloxamers are nonionic triblockcopolymers composed of a central hydrophobic chain of polyoxypropylene(poly(propylene oxide)) flanked by two hydrophilic chains ofpolyoxyethylene (poly(ethylene oxide)). Certain poloxamers, such asthose listed herein, are also known by the trade names Pluronic®available from suppliers such as BASF AG (Ludwigshafen, Germany).Because the lengths of the polymer blocks can be customized, manydifferent poloxamers exist that have slightly different properties.Further exemplary Pluronic® poloxamers include, but are not limited toPluronic® 10R5, Pluronic® 17R2, Pluronic® 17R4, Pluronic® 25R2,Pluronic® 25R4, Pluronic® 31R1, Pluronic® F 108 Cast Solid Surfacta,Pluronic® F 108 NF, Pluronic® F 108 Pastille, Pluronic® F 108 Prill,Pluronic® F 108NF Prill Poloxamer 338, Pluronic® F 127, Pluronic® F 127Prill, Pluronic® F 127 NF, Pluronic® F 127 NF 500 BHT Prill, Pluronic® F127 NF Prill Poloxamer 407, Pluronic® F 38, Pluronic® F 38 Pastille,Pluronic® F 68, Pluronic® F 68 Pastille, Pluronic® F 68 LF Pastille,Pluronic® F 68 NF, Pluronic® F 68 NF Prill Poloxamer 188, Pluronic® F 68Prill, Pluronic® F 68 Prill, Pluronic® F 77, Pluronic® F 77Micropastille, Pluronic® F 87, Pluronic® F 87 NF, Pluronic® F 87 NFPrill Poloxamer 237, Pluronic® F 87 Prill, Pluronic® F 88, Pluronic® F88 Pastille, Pluronic® F 88 Prill, Pluronic® F 98, Pluronic® F 88 Prill,Pluronic® F 98, Pluronic® F 98 Prill, Pluronic® L 10, Pluronic® L 101,Pluronic® L 121, Pluronic® L 31, Pluronic® L 35, Pluronic® L 43,Pluronic® L 44, Pluronic® L 61, Pluronic® L 62, Pluronic® L 62 LF,Pluronic® L 62D, Pluronic® L 64, Pluronic® L 81, Pluronic® L 92,Pluronic® L44 NF INH surfactant Poloxamer 124, Pluronic® N 3, Pluronic®P 103, Pluronic® P 104, Pluronic® P 105, Pluronic® P 123 Surfactant,Pluronic® P 65, Pluronic® P 84, Pluronic® P 85, or combinations thereof.

In certain embodiments, the at least one surface active agent comprisesa polysorbate present from, for example, 15% (wt/wt) to 35% (wt/wt) ofthe composition.

In certain embodiments, the polysorbate is present at, for example, 15%,16%, 17%, 18%, 19%, 20%, 21%, 22%, 23%, 24%, 25%, 26%, 27%, 28%, 29%,30%, 31%, 32%, 33%, 34%, or 35% (wt/wt) of said composition.

In certain embodiments, the polysorbate is present at, for example, 31%(wt/wt) of the composition.

In certain embodiments, the polysorbate is polysorbate 20, polysorbate60, polysorbate 80 or a combination thereof. Typically, but notnecessarily, the polysorbate is polysorbate 80.

In certain other embodiments, the at least one surface active agentcomprises a poloxamer present from, for example, 0.1% (wt/wt) to 5%(wt/wt) of the composition.

In certain embodiments, the poloxamer is present at, for example, 0.1%,0.2%, 0.3%, 0.4%, 0.5%, 0.6%, 0.7%, 0.8%, 0.9%, 1%, 1.5%, 2%, 2.5%, 3%,3.5%, 4%, 4.5%, or 5% (wt/wt) of said composition.

In certain embodiments, the poloxamer is present at, for example, 0.7%(wt/wt) of the composition.

In certain embodiments, the poloxamer is Pluronic® F87[(HO(C₂H₄O)₆₄(C₃H₆O)₃₇(C₂H₄O)₆₄H], which is poloxamer 237.

In certain embodiments, the compositions described herein comprise acombination of polysorbate 80 and the poloxamer Pluronic® F87[(HO(C₂H₄O)₆₄(C₃H₆O)₃₇(C₂H₄O)₆₄H], which is poloxamer 237.

In certain embodiments, the at least one surface active agent comprisesa combination of nonionic surfactants. For example, the combination ofnonionic surfactants can comprise a combination of a polysorbate and apoloxamer totaling at least, for example, 30% (wt/wt) of saidcomposition. In certain embodiments, the polysorbate is polysorbate 80and the poloxamer is poloxamer 237 (Pluronic® F87). In certainembodiments, the polysorbate 80 is present at, about 31% (wt/wt) and thepoloxamer 237 is present at, about 0.7% (wt/wt) of said composition.

In some embodiments, the compositions include one or more anionicsurface active agents. Exemplary “anionic surface active agents”include, but are not limited to, N-acyl-L-glutamic acid diethanolamine,N-acyl-L-glutamic acid triethanolamine, sodium N-acyl-L-glutamate,sodium alkanesulfonate, ammonium alkyl (C12, C14, C16) sulfate, alkyl(C11, C13, C15) sulfuric acid triethanolamine, alkyl (C11, C13, C15)sulfuric acid triethanolamine, alkyl (C12 to C14) sulfuric acidtriethanolamine, liquid alkylsulfuric acid triethanolamine, sodium alkyl(C12, C13) sulfate, liquid sodium alkylsulfate, sodium isoethionate,sodium lacto-isostearate, disodium undecylenoylamido ethylsulfosuccinate, triethanolamine sulfooleate, sodium sulfooleate,disodium oleamide sulfosuccinate, potassium oleate, sodium oleate,morpholine oleate, oleoyl sarcosine, oleoyl methyltaurine sodium salt,potassium-containing soap base, liquid base for potassium soap,potassium soap, carboxylated polyoxyethylene tridodecyl ether, sodiumsalt (3 ethyle oxide “E.O.”) of carboxylated polyoxyethylene tridodecylether, triethanolamine N-hydrogenated tallow fatty-acyl-L-glutamate,sodium N-hydrogenated tallow fatty-acyl-L-glutamate, sodium hydrogenatedcoconut fatty acid glyceryl sulfate, sodium diundecylenoylamido ethylsulfosuccinate, sodium stearyl sulfate, potassium stearate,triethanolamine stearate, sodium stearate, sodiumN-stearoyl-L-glutamate, disodium stearoyl-L-glutamate, stearoylmethyltaurine sodium salt, sodium dioctyl sulfosuccinate, liquid sodiumdioctyl sulfosuccinate, liquid disodium polyoxyethylene monooleylamidosulfosuccinate (2 E.O.), disodium polyoxyethylene lauroyl ethanolamidesulfosuccinate (5 E.O.), disodium lauryl sulfosuccinate, diethanolamidecetyl sulfate, sodium cetyl sulfate, soap base, sodium cetostearylsulfate, triethanolamine tridecyl sulfate, potassium palmitate, sodiumpalmitate, palmitoyl methyltaurine sodium salt, liquid castor oil fattyacid sodium salt (30%), ammonium polyoxyethylene alkyl ether sulfate (3E.O.), liquid diethanolamine polyoxyethylene alkyl (C12, C13) ethersulfate, liquid triethanolamine polyoxyethylene alkyl ether sulfate (3E.O.), triethanolamine polyoxyethylene alkyl (C11, C13, C15) ethersulfate (1 E.O.), triethanolamine polyoxyethylene alkyl (C12, C13) ethersulfate (3 E.O.), liquid sodium polyoxyethylene alkyl ether sulfate (3E.O.), sodium polyoxyethylene alkyl (C11, C13, C15) ether sulfate (1E.O.), sodium polyoxyethylene alkyl (C11 to C15) ether sulfate (3 E.O.),sodium polyoxyethylene alkyl (C12, C13) ether sulfate (3 E.O.), sodiumpolyoxyethylene alkyl (C12 to C14) ether sulfate (3 E.O.), sodiumpolyoxyethylene alkyl (C12 to C15) ether sulfate (3 E.O.), disodiumpolyoxyethylene alkyl (C12 to C14) sulfosuccinate (7 E.O.), sodiumpolyoxyethylene undecyl ether sulfate, liquid sodium polyoxyethyleneoctyl phenyl ether sulfate, ammonium polyoxyethylene oleyl ethersulfate, disodium polyoxyethylene lauryl sulfosuccinate, sodiumpolyoxyethylene nonyl phenyl ether sulfate, sodium polyoxyethylenepentadecyl ether sulfate, triethanolamine polyoxyethylene myristyl ethersulfate, sodium polyoxyethylene myristyl ether sulfate, sodiumpolyoxyethylene myristyl ether sulfate (3 E.O.), liquid sodiumpolyoxyethylene lauryl ether acetate (16 E.O.), ammonium polyoxyethylenelauryl ether sulfate (2 E.O.), triethanolamine polyoxyethylene laurylether sulfate, sodium polyoxyethylene lauryl ether sulfate,diethanolamine myristyl sulfate, sodium myristyl sulfate, potassiummyristyl sulfate, sodium N-myristoyl-L-glutamate, sodiummyristoylmethylaminoacetate, liquid myristoyl methyl-alanine sodiumsalt, myristoyl methyltaurine sodium salt, medicinal soaps,triethanolamine/magnesium coco alkyl sulfate, triethanolamine N-coconutoil fatty-acyl-L-glutamate, sodium N-coconut oil fatty-acyl-L-glutamate,sodium coconut oil fatty acid ethyl ester sulfonate, coconut oil fattyacid potassium salt, liquid coconut oil fatty acid potassium salt,sodium N-coconut oil fatty/hydrogenated fatty-acyl-L-glutamate, coconutoil fatty acid sarcosine, coconut oil fatty acid sarcosinetriethanolamine salt, coconut oil fatty acid sarcosine sodium salt,coconut oil fatty acid triethanolamine salt, liquid triethanolamine saltof coconut oil fatty acid, coconut oil fatty acid sodium salt, coconutoil fatty acid methyl alanine sodium salt, liquid coconut oil fatty acidmethyl alanine sodium salt, coconut oil fatty acid methyltaurinepotassium salt, coconut oil fatty acid methyltaurine sodium salt, sodiumlaurylamino dipropionate, liquid sodium laurylamino dipropionate (30%),sodium lauryl sulfoacetate; sodium lauryl benzenesulfonate, laurylsulfate, ammonium lauryl sulate, potassium lauryl sulfate,diethanolamine lauryl sulfate, triethanolamine lauryl sulfate, sodiumlauryl sulfate, magnesium lauryl sulfate, monoethanolainine laurylsulfate, potassium laurate, lauric acid triethanolamine, liquid lauricacid triethanolamine, sodium laurate, lauric acid/myristic acidtriethanolamine, lauroyl-L-glutamic acid triethanolamine, sodiumN-lauroyl-L-glutamate, lauroyl sarcosine, lauroyl sarcosine potassium,liquid lauroyl sarcosine triethanolamine salt, lauroyl sarcosine sodium,liquid lauroyl methyl-.beta.-alanine sodium salt, lauroyl methyltaurinesodium salt, liquid lauroyl methyltaurine sodium salt, or combinationsthereof.

In certain embodiments, said anionic surfactant(s) comprise from, forexample, about 0.05% (wt/wt) to about 25% (wt/wt) of said composition.In certain embodiments, said anionic surfactant(s) comprise from, forexample, about 0.05% (wt/wt) to about 15% (wt/wt) of said composition.In certain embodiments, said anionic surfactant(s) comprise, forexample, 0.05% (wt/wt) to 5% (wt/wt) of said composition. In certainembodiments, said anionic surfactant(s) comprise, for example, 0.5%(wt/wt) to 3% (wt/wt) of said composition. In certain embodiments, saidanionic surfactant(s) comprise, for example, 0.7% (wt/wt) of saidcomposition. In certain embodiments, said anionic surfactant(s) comprisesodium lauryl sulfate.

In some embodiments, the compositions include additional surface activeagents such as the zwitterionic and cationic surface active agents.Examples of such surface active agents include, but are not limited tothe bile acids (e.g., cholic acid, chenodeoxycholic acid, glycocholicacid, glycodeoxycholic acid, taurocholic acid, taurochenodeoxycholicacid, taurolithocholic acid, deoxycholic acid, lithocholic acid, andursodeoxycholic acid and salts thereof, e.g., sodium, potassium,lithium), natural emulsifiers (e.g. acacia, agar, alginic acid, sodiumalginate, tragacanth, chondrux, cholesterol, xanthan, pectin, gelatin,egg yolk, casein, wool fat, cholesterol, wax, and lecithin), long chainamino acid derivatives, high molecular weight alcohols (e.g. stearylalcohol, cetyl alcohol, oleyl alcohol, triacetin monostearate, ethyleneglycol distearate, glyceryl monostearate, and propylene glycolmonostearate, polyvinyl alcohol), carbomers (e.g. carboxy polymethylene,polyacrylic acid, acrylic acid polymer, and carboxyvinyl polymer),carrageenan, cellulosic derivatives (e.g. carboxymethylcellulose sodium,powdered cellulose, hydroxymethyl cellulose, hydroxypropyl cellulose,hydroxypropyl methylcellulose, methyl cellulose), polyoxyethylene esters(e.g. polyoxyethylene monostearate [Myrj 45], polyoxyethylenehydrogenated castor oil, polyethoxylated castor oil, polyoxymethylenestearate, and Solutol), sucrose fatty acid esters, polyethylene glycolfatty acid esters (e.g. Cremophor), polyoxyethylene ethers, (e.g.polyoxyethylene lauryl ether [Brij 30]), poly(vinyl-pyrrolidone),diethylene glycol monolaurate, triethanolamine oleate, sodium oleate,potassium oleate, ethyl oleate, oleic acid, ethyllaurate, sodium laurylsulfate, cetrimonium bromide, cetylpyridinium chloride, benzalkoniumchloride, docusate sodium or combinations thereof.

Without being bound to any particular theory, it is believed that thecompositions described herein will increase fatty acid absorption inpatients with fat malabsorption syndromes and other disease states asdescribed herein. This is because the compositions described hereincomprising the omega-3 fatty acid(s) and the at least one surface activeagent self-micellize in a manner similar to that in the body. Whenmammals eat, micelles are naturally formed around the fats ingested.These natural micelles form in the presence of bile regardless of the pHin the stomach or intestines. Natural micelles form around fats anddigestive enzymes, entrapping both. Within the micelles, the enzymesquickly cleave the triglycerides or fatty acid esters into free fattyacids and monoglycerides. Natural micelles fuse with the cell membranewhen they come in contact with the brush barrier of the intestinal wall,delivering their cargo of free fatty acids and monoglycerides, amongother components and compounds, into the cells lining the intestine andare absorbed. Natural micelles that form around omega-3 fatty acids havetwo characteristics that limit their effectiveness. Firstly, given theformation of natural micelles require the food-dependent release of bilesalts, natural micelles form only when food is present. Secondly, sincenatural micelles require fats from the consumed food in order to formthe outer shell of the micelle, the amount of lipids transported intothe intestinal wall is directly correlated with the amount of fatco-consumption. As a result, unless a lipophilic compound isco-administered with food, very few natural micelles form and, unlessthat food contains a sufficient amount of fat, very few natural micellesform. It then follows that the lower the natural micelle formation, theless efficient the natural process is at delivering said lipophiliccompound, among other components and compounds, in micellar form, to theintestinal lumen.

In contrast to natural micelles, the formation of artificial micelleshas hitherto been very sensitive to the pH of the liquid in which theyare being created and to the presence of other substances in thatliquid. In addition, artificial micelles are in general either fragileand break easily or are too rigid and never release their cargo, makingit difficult for them to deliver adequate amounts of free fatty acidsand monoglycerides to the intestinal lining. The micelles formed by thecompositions described herein, however, overcome the deficiencies ofboth artificial and natural human micelles in the delivery of omega-3fatty acids, while at the same time mimicking some of thecharacteristics of natural human micelles.

“Emulsion” is a very broad term describing particles that may beamorphous, liquid-crystalline, bilayer, monolayer or any mixture ofthese structures. An emulsion is a more general term that describes adispersed phase in a continuous phase that is commonly seen as oil inwater (o/w), water in oil (w/o) or even more complex mixtures such aswater in oil in water (w/o/w). Emulsion particle sizes can vary widelyfrom 10 nanometers to 100 micrometers.

Solutions in lipid vehicles containing surfactants, which constituteself-emulsifying drug delivery systems (SEDDS), and which effectspontaneous emulsification upon contact of the lipid with fluids in thegastrointestinal tract (GI tract), are known in the art, and areclassified as “self-emulsified”, “self-micro”, and “self-nano”, drugdelivery systems (herein referred to as “SEDDS,” “SMEDDS,” and “SNEDDS,”respectively). The SEDDS, SMEDDS, and SNEDDS are emulsions. Theseemulsions have the sole purpose of dispersing lipids, such as oils, intosmall droplets in order to increase the surface area available tointeract with the body's natural bile salts, pancreatic lipases,co-lipase, and phospholipase A2.

SEDDS are formulated with mixtures of lipid vehicles, non-ionicsurfactants and a drug in the absence of water, and are assumed to existas transparent isotropic solutions. These systems have a uniqueproperty—they are able to self-emulsify rapidly in the gastrointestinalfluids, forming fine oil-in-water emulsions (droplet size diameter <300nm) under gentle agitation provided by gastrointestinal motion. SEDDSare commonly suitable for oral delivery in soft and hard gelatin or hardhydroxypropyl methylcellulose (HPMC) capsules.

SMEDDS are defined as isotropic mixtures of an oil, surfactant,co-surfactant (or solubilizer) and a drug. Such systems form fineoil-in-water microemulsions under gentle agitation provided by digestivemotility of the stomach and intestine following dilution by the aqueousphase in vivo. SMEDDS are distinguished from SEDDS by smaller emulsiondroplets produced on dilution, resulting in a transparent or translucentstable dispersion. Mean droplet size after dilution is <100 nm in thecase of SMEDDS or <300 nm in the case of SEDDS. SMEDDS generally containrelatively high concentrations of surfactant (typically 30 to 60%, m/m),and optionally also hydrophilic co-solvents (e.g., propylene glycol,polyethylene glycols). They are often described as microemulsionpre-concentrates because the microemulsion is formed on dilution inaqueous media.

SNEDDS are defined as self-nanoemulsifying drug delivery systems thatspontaneously form transparent to opalescent oil-in-water dispersions ofapproximately 200 nm in size upon dilution with water under gentlestirring.

As these various emulsions move through the GI tract various processesoccur. First the components are dispersed to form lipid droplets, oremulsion droplets. This is followed by lipolysis and solubilization ofthe digestion products by bile acids, which may then form colloidalmixed micelles. It is believed that the drug then partitions from theemulsion oil droplets and bile salt mixed micelles to be absorbed by themucosal cells of the intestinal wall.

Thus, because SEDDS, SMEDDS, and SNEDDS leverage the body's naturalprocess to be efficient, they also necessarily rely on the naturalprocesses to be effective. This is particularly important because inorder to form micelles, SEDDS, SMEDDS, and SNEDDS require the presenceof bile salts. Since bile salts are emitted into the GI tract in mammalsas a response to food consumption, then it follows that SEDDS SMEDDS,and SNEDDS are not able to use bile salts to form micelles in the GItract unless food intake has induced the introduction of bile salts intothe GI tract.

On the contrary, the present invention is directed toward aSelf-Micellizing Drug Delivery System (SMDDS). This system isdifferentiated from the SEDDS, SMEDDS and SNEDDS as described above, inthat the particularly described combination of surface active agent(polysorbate and poloxamer 237) in the relative amounts and overallconcentrations as herein described, are unique in their ability todirectly and spontaneously form stable micelles in a size range of about1 to about 10 micron without relying on the presence of bile salts. Thisrepresents a distinct and unique advantage of the present invention, inthat, unlike SEDDS, SMEDDS, and SNEDDS, the present invention is able todeliver lipids through the intestinal wall by way of a plurality ofspontaneously formed stable micelles. Unlike SEDDS, SMEDDS, and SNEDDS,which function to disperse lipids to increase surface area, the presentinvention encapsulates the lipids with surface active agents, thusbringing the lipids together, whereby predictable and repeatableenhancements in bioavailability are realized, absent any food effect.

As opposed to an emulsion, a micelle is a particle of colloidaldimensions that exists in equilibrium with molecules or ions in asolution from which it is formed. Micelles are a particular type ofparticle (oil particle in water) which form a structure wherein thehydrophilic component is external and the hydrophobic component isinternal. More specifically, micelles are usually formed from singlechain lipids and surfactants, and a micelle is always a monolayerparticle with the hydrophilic head facing the aqueous phase and thehydrophobic tail facing the oil phase.

One method of determining a human's normal weight range is to assess thesubject's Basal Metabolic Rate (BMR), which takes into account asubject's height, weight, age and gender. BMR determinations are wellknown in the art. Accordingly, it is believed, without being bound toany particular theory, that the compositions or a plurality of micellespre-formed therefrom described herein will lower a human's weight, ifthe human is overweight, to within the normal range as determined by thehuman's BMR. Conversely, it is believed, without being bound to anyparticular theory, that the compositions or a plurality of micellespre-formed therefrom described herein will increase a human's weight, ifthe human is underweight, malnourished, or is suffering from amalabsorption syndrome, to within the normal range as determined by thehuman's BMR.

It is known in the art that food intake in a subject having a normalweight stimulates adipose tissue to increase leptin levels. Leptin is ahormone that tells the brain it is full and increases metabolism. Thus,in starvation mode there is a decrease in leptin, telling the brain itis hungry, which in turn stimulates food intake and decreases energyexpenditure. In an obese subject, however, the brain is resistant toleptin signaling despite leptin levels being high, thus, the subjectbelieves it is not satiated and continues eating and gaining weight.

Omega-3 fatty acids are known to decrease circulating leptin levels.Accordingly, without being bound to any particular theory, it isbelieved that the enhanced absorption of omega-3 fatty acids provided bythe compositions or a plurality of micelles pre-formed therefromdescribed herein will regulate leptin levels and signal a subject'sbrain accordingly to either decrease leptin levels if the subject isoverweight or increase leptin levels if the subject is underweight,malnourished, or is suffering from a malabsorption syndrome. Thisincrease or decrease in metabolism, as the case may be, is reflected byan increase or decrease in circulating free fatty acids when compared tocirculating free fatty acid levels at the start of the dosing regimen.

Without being bound to any particular theory, it is believed that theself-micellizing compositions described herein will regulate metabolismin a subject administered said compositions to facilitate returning thesubject's weight to within a normal range for that particular subject.

Accordingly, in at least one embodiment of the present invention, amethod is disclosed for administering self-micellizing compositionsdescribed herein to facilitate a subject's weight gain, loss, or returnto normal weight, or combinations thereof over time.

In at least one embodiment of the present invention, a composition isdisclosed which can be used to increase the weight of a mammal, such aslivestock.

Also disclosed herein in at least one embodiment of the invention is amethod for administering a composition as provided herein to increasethe weight of a mammal, such as livestock.

In at least one embodiment, the compositions described herein comprisingthe omega-3 fatty acid(s) and the at least one surface active agent, forexample the combination of polysorbate 80 and poloxamer 237,spontaneously form micelles when encountering aqueous liquids. Thesemicelles form regardless of the pH or the nature and concentration ofother suspended materials in the liquid, do not require bile to form,and form micelles that remain stable almost indefinitely. The micellesform whether or not food is present or whether food that is present ishigh or low in fat. Like human micelles, the micelles formed by thecompositions described herein form around omega-3 fatty acids and entrapboth fats and enzymes, allowing rapid digestion and the formation offree fatty acids and monoglycerides. Without being held to any onetheory, it is believed that the micelles formed by the compositionsdescribed herein are similar in size to human micelles and share theability to rupture at the intestinal brush barrier. Without being heldto any one theory, it is believed that either by friction or a chemicalreaction, the micelle ruptures and spills its contents onto the liningof the intestines and the intestines absorb the omega-3 fatty acids.

Accordingly, it is believed that compositions described herein canovercome one of the primary challenges of malabsorption syndromes, suchas SBS i.e., the delivery of adequate amount of free fatty acids to thebloodstream of patients suffering from the hepatic complications of SBSwithout exacerbating liver dysfunction, as does TPN treatment, ortriggering fat malabsorption or dumping. This would also allow for moreconsistent dosing and thus produce more reliable results.

In certain embodiments, the compositions described herein self-micellizein an aqueous medium. The aqueous medium can include, for example, 0.1NHCl. It is well accepted that 0.1N HCl (simulated gastric fluid) servesas a proxy for the acidity of stomach contents. Accordingly, and withoutbeing bound by theory, it is believed that the compositions describedherein can self-micellize in situ in the stomach or small intestine. Incertain embodiments, the compositions described herein more efficientlyand effectively deliver omega-3 fatty acid esters through the intestinaltract when administered with or without food.

In addition to forming micelles in situ, in other embodiments,compositions comprising micelles are provided, wherein the micelles areformed by the addition of an aqueous medium to a composition of any oneof the embodiments provided herein prior to administration of saidcomposition to a subject in need of treatment. Alternatively, micellescan also be formed when the compositions are added to an aqueous medium.In certain embodiments, the micelles have a diameter of up to 10 μm. Inother embodiments, substantially all of the micelles have an averagediameter of from 1 μm to 10 μm. In certain embodiments, the micelleshave an average diameter of, for example, 1, 2, 3, 4, 5, 6, 7, 8, 9 or10 μm. In certain embodiments, said micelles are stable at ambienttemperature and in certain other embodiments, said micelles are stableat about normal mammalian body temperatures.

The surface active agents comprising the compositions suitable forself-micellization as described herein generally have an HLB from, forexample, 12 to 18. In certain embodiments, said surface active agentshave an HLB from, for example, 12.0 to 14.0. In certain embodiments,said surface active agents have an HLB from, for example, 13.0 to 14.0.In certain embodiments, said surface active agents have an HLB from, forexample, 13.5 to 13.8. The total HLB of all the surface active agents orsurfactants used in the composition is generally from, for example, 12to 18. In some embodiments, the total HLB of all surface active agentsused in the composition is generally from, for example, 12 to 15. Insome embodiments, the total HLB of all surface active agents orsurfactants used in the composition is generally from, for example, 13to 15.

In certain embodiments, the at least one surface active agent orsurfactant has an HLB of at least, for example, 8.0. In someembodiments, said surface active agent(s) or surfactant(s) have acombined HLB in the range of from, for example, 13 to 15. As the HLBvalue of the surface active agent(s) or surfactant(s) increases, theamount of surface active agent or surfactant needs to be decreased.

The compositions described herein can further comprise at least oneantioxidant. The antioxidant(s) suitable for use in the instant omega-3fatty acid compositions, include, but are not limited to tocopherolsand/or tocotrienols and can be present from, for example, 0.01% to 5%(wt/wt) of the composition. In certain such embodiments, the tocopherolsand/or tocotrienols can be present at, for example, 0.01%, 0.05%, 0.1%,0.2%, 0.3%, 0.4%, 0.5%, 0.6%, 0.7%, 0.8%, 0.9%, 1%, 1.5%, 2%, 2.5%, 3%,3.5%, 4%, 4.5% or 5% by weight of the compositions. In certain suchembodiments, the antioxidant is α-tocopherol present at, for example, 2%by weight of the composition.

In certain embodiments, the composition further comprises a terpene. Incertain embodiments, the terpene is d-limonene. In one embodiment, theterpene is a cyclic terpene. In one embodiment, the terpene isd-limonene ((+)-limonene), which is the (R)-enantiomer. In oneembodiment, the terpene is L-limonene, which is the (S)-enantiomer. Inone embodiment, the terpene is racemic limonene, known as dipentene. Inanother embodiment, the terpene is a terpenoid. In another embodiment,the terpene or terpenes are derived from a natural oil (e.g., a citrusoil such as orange oil). Other terpenes are contemplated, such asmonoterpenes (e.g., terpinenes, terpinolenes, phellandrenes, ormenthol), having structures that are similar to d-limonene. In certainembodiments, the compositions further comprise substantially pured-limonene from, for example, 0.1% to 5% by weight of the composition.In certain other embodiments, the compositions further comprise naturalorange oil from, for example, 0.1% to 5% by weight of the composition.Compositions comprising d-limonene or orange oil can aid in theelimination and/or minimization of side effects from the oraladministration of the instant omega-3 fatty acid compositions. Such sideeffects include regurgitation, frequency of belching, gastroesophagealreflux disease (GERD), bloating, increased intestinal gas, fish taste,fishy breath, fish smell, nausea, diarrhea, or combinations thereof.

Methods for Treating Age-Related Macular Degeneration

Methods are provided of treating AMD in a subject in need of treatment,which method comprises administering to said subject a therapeuticallyeffective amount of a composition of any one of the embodiments providedherein, or a pre-formed micelles according to the respective embodimentsdescribed herein. The compositions described herein can be administeredin conjunction with other medications and/or nutritional supplementsprescribed for treating AMD. In certain embodiments, non-limitingexamples of non-omega-3 fatty acid nutritional supplements include, forexample, a combination of vitamin C, vitamin E, beta-carotene, zinc,copper, magnesium, manganese, calcium, vitamin A, vitamin D, vitamin K2,lutein and zeaxanthin. Non-limiting examples of non-omega-3 fatty acidactive agents include, for example, verteporfin (VISUDYNE); theantioxidant carotenoids crocin and crocetin, as found in, for example,Saffron (Crocus sativus), and inhibitors of angiogenesis, such as forexample, BEVACIZUMAB (AVASTIN®), RANIBIZUMAB (LUCENTIS®), PEGAPTANIB(MACUGEN®) AND AFLIBERCEPT (EYLEA®).

The compositions described herein can also be administered for treatingJuvenile Macular Degeneration, which includes Stargardt's disease, Bestdisease, and juvenile retinoschisis by administering the compositionsdescribed herein to a human juvenile.

In certain embodiments, the amount of the non-omega-3 fatty acid activeagents administered in conjunction with the compositions describedherein can be reduced to avoid or minimize any side effects resultingfrom the non-omega-3 fatty acid active agents when such agents areadministered alone at higher concentrations.

In at least one embodiment, a method is provided for treating AMD byadministering, to a patient in need thereof, a therapeutically effectiveamount of a composition comprising omega-3 DPA fatty acid (either in thetriglyceride, ester or free fatty acid ester form) and at least onesurface active agent in an amount and a combination effective to causesaid compositions to spontaneously self-micellize when in contact withan aqueous medium, thereby forming a plurality of stable micelles havinga particle size within a range of about 1 μm to about 10 μm.

Methods for Treating Fat Malabsorption Syndromes or Disorders

Methods are provided for treating one or more fat malabsorption syndromeor disorder in a subject in need of treatment, which method comprisesadministering to said subject a therapeutically effective amount of acomposition of any one of the embodiments provided herein, or apre-formed micelle mixture of any one of the embodiments providedherein.

Accordingly, in certain embodiments, the fat malabsorption syndrome canbe the result of a disorder in the intestinal processes of digestionand/or transport of nutrients across the intestinal mucosa into thesystemic circulation. The fat malabsorption syndrome can be either acongenital abnormality in the digestive or absorptive processes or asecondarily acquired disorder resulting from a disorder of theintestinal processes of digestion and/or transport of nutrients acrossthe intestinal mucosa into the systemic circulation.

Exocrine pancreatic insufficiency can also result in a fat malabsorptionsyndrome or disorder. Exocrine pancreatic insufficiency can be theresult of pancreatitis, pancreatic cancer, pancreatic resection, cysticfibrosis, Shwachman-Diamond syndrome, Johnson-Blizzard syndrome, andPearson syndrome. Accordingly, in certain embodiments, the compositionsdescribed herein can be used for the treatment of fat malabsorptionsyndrome resulting from exocrine pancreatic insufficiency.

Obstructive biliary or cholestatic liver disease or extensive intestinalmucosal disease, such as that which occurs in celiac disease can resultin fat malabsorption syndromes. Thus, in certain embodiments, thecompositions described herein can be administered to a human in need ofsuch administration for the treatment of fat malabsorption resultingfrom obstructive biliary or cholestatic liver disease or extensiveintestinal mucosal disease.

Fat malabsorption syndromes can be the result of functional or anatomicloss of extensive segments of small intestine leading to a severedecrease in intestinal absorptive capacity, often referred to asShort-Bowel Syndrome (SBS), which may be congenital or acquired as aresult of surgery due to Crohn's Disease or necrotizing enterocolitis ininfants or neonates. In certain embodiments, the administration of thecompositions described herein to a human, particularly a neonate, inneed of such administration, can treat fat malabsorption syndromesresulting from SBS or Crohn's Disease.

In order to increase the absorption of dietary fats, includingdocosahexaenoic acid, through enteral administration, a combination offatty acids and surface active agents, specifically poloxamers andpolysorbates, can be combined to form self-micellizing micelles.Attempts to create artificial micelles have generally resulted inartificial micelles that are fragile and break easily, making itdifficult for the particles to deliver adequate amounts of free fattyacids and monoglycerides to the intestinal lining for absorption. Forthis reason, creating micelles mimicking necessary characteristics ofnatural human micelles is important, especially having a size of aboutfive microns in diameter to carry its cargo of dietary fats (or otherlipophilic materials) to and through the intestinal cell wall is a novelmeans of creating bioavailable dietary fats for the treatment ofmalabsorption.

In one exemplary embodiment of the present invention, a novelformulation was utilized containing docosahexaenoic acid in combinationwith a blend of surface active agents including polysorbates andpoloxamers, and a study was carried out in a porcine model of SBS. Themethodology and results of this experiment are set forth in Example 3.

It was concluded that enteral administration of a novel DHA preparationthat forms micelles independent of bile salts results in increased fattyacid absorption, weight gain and improved intestinal morphologicadaptation in a neonatal porcine model of SBS.

In certain embodiments, administration of the compositions describedherein can be administered in conjunction with a non-omega-3 fatty acidactive agent for the treatment of SBS. Non-limiting examples of suchnon-omega-3 fatty acid active agents can include L-GLUTAMINE(NUTRESTORE®), RECOMBINANT SOMATOTROPIN (ZORBTIVE®), AND TEDUGLUTIDE(GATTEX®). Other such non-omega-3 fatty acid active agents may includeconjugated bile acids or opium tincture. In certain embodiments, theamount of the non-omega-3 fatty acid active agent(s) administered inconjunction with the compositions described herein can be reduced toavoid or minimize any side effects resulting from the non-omega-3 fattyacid active agents when such agents are administered alone at higherconcentrations.

Methods for Treating NAFLD and/or NASH

Methods are provided of treating NAFLD and/or NASH in a subject in needof treatment, which method comprises administering to said subject atherapeutically effective amount of a composition of any one of theembodiments provided herein, or pre-formed micelles according to therespective embodiments described herein.

The compositions described herein can be administered concomitantly withother non-omega-3 fatty acid medications prescribed for treating NAFLDand/or NASH. Non-limiting examples of such non-omega-3 fatty medicationscan include, lipid lowering or cholesterol lowering agents selected fromthe group consisting of cholesterol absorption inhibitors, bile acidsequestrants/resins, statins, niacin and derivatives, MTP inhibitors,fibrates and CETP inhibitors, insulin sensitizers, hypolipidemics,anti-inflammatory medications, and thiazolidinediones. In certainembodiments, the amount of the non-omega-3 fatty acid active agent(s)administered in conjunction with the compositions described herein canbe reduced to avoid or minimize any side effects resulting from thenon-omega-3 fatty acid active agents when such agents are administeredalone at higher concentrations.

In certain embodiments, the compositions, or a plurality of micellespre-formed therefrom, described herein, provide for a method of treatinga Non-Alcoholic Fatty acid Liver Disease (NAFLD) comprising the step ofadministering to a subject in need of such administration thecompositions or a plurality of micelles pre-formed therefrom describedherein.

In certain embodiments, the compositions, or a plurality of micellespre-formed therefrom, described herein, provide for a method of treatingNon-Alcoholic Steatohepatitis (NASH) comprising the step ofadministering to a subject in need of such administration thecompositions or a plurality of micelles pre-formed therefrom describedherein.

In certain embodiments, the compositions, or a plurality of micellespre-formed therefrom, described herein, provide for a reduction in bodyweight (or weight loss) in a subject diagnosed with NAFLD and/or NASH.

In certain embodiments, the compositions, or a plurality of micellespre-formed therefrom, described herein, provide for a method of reducingliver weight in a subject diagnosed with NASH.

In certain embodiments, the compositions, or a plurality of micellespre-formed therefrom, described herein, provide for a method of reducingwhole blood glucose in a subject diagnosed with NASH.

In certain embodiments, the compositions, or a plurality of micellespre-formed therefrom, described herein, provide for a method of reducingliver triglyceride levels in a subject diagnosed with NAFLD and/or NASH.

In certain embodiments, the compositions, or a plurality of micellespre-formed therefrom, described herein, provide for a method of reducingdevelopment of liver fibrosis in a subject diagnosed with NAFLD and/orNASH.

A nine-week preclinical study of certain embodiments of the presentinvention was conducted using a mouse model which progresses from NAFLDto NASH between 5 to 9 weeks of age. Within the disease progression ofthis animal model, the liver triglycerides reach a point around week 6where the inflammation has progressed substantially over week 5. Thestudy contained three groups, a control group given a placebo from agefive to nine weeks, and two trial groups, a first trial groupadministered a composition disclosed herein for 4 weeks, from five tonine weeks of age, and a second trial group administered the same saidcomposition disclosed herein for 3 weeks, from six to nine weeks of age.

The study showed that animals for which treatment was started later, the3 week treatment group, had significantly reduced liver triglyceridecontents compared with the control group. Although the livertriglycerides tended to decrease in both treatment groups, the 4 weektreated group (from five to nine weeks) showed a non-significant change(at 0.05) and the group treated for 3 weeks (from six to nine weeks)showed a statistically significant decrease, as illustrated in FIG. 3.This 3 week treatment group also showed a tendency for reduced wholeblood glucose levels compared with the control group. The significantdecrease in the liver triglycerides in the more progressed groupdemonstrate that certain embodiments of the present invention canimprove lipid and glucose metabolism in the liver. These results alsodemonstrate that omega-3 fatty acids can improve lipid and glucosemetabolism in the liver.

The effect of the treatment was not merely through an induction ofweight loss by the animals. In fact, the study found that body weightgradually increased during the treatment period in all groups and thatthere was no difference in mean body weight between groups. Although theend liver weight of the subjects in the treatment groups was lower thanthat of the subjects in the control group, the difference was notstatistically significant. In addition, both treatment groups trended toreduce alpha-SMA (a marker of myofibroblast associated with thedevelopment of liver fibrosis) mRNA expressions in the liver, a keyindicator of the treatment's demonstrated ability to delay theprogression to NASH. Without being held to any one theory, we believethe study demonstrates the ability of at least one composition disclosedherein to deliver polyunsaturated EPA and DHA fatty acids to mammalswith NAFLD for incorporation into the cell membrane of erythrocytes,wherein said delivery can reduce the triglyceride level in the liver andthereby slow the disease's progression to NASH. The study may alsoindicate that administration of compositions disclosed herein mightprovide a larger medicinal response when provided later in the diseaseprogression from NAFLD to NASH.

Methods for Treating Neurodegenerative Diseases

Methods are provided of treating neurodegenerative disease in a subjectin need of treatment, which method comprises administering to saidsubject a therapeutically effective amount of a composition of any oneof the embodiments provided herein, or pre-formed micelles according tothe respective embodiments described herein. Non-limiting examples ofneurodegenerative diseases that can be treated by the compositionsdescribed herein include PD, AD, MS, Epilepsy, and ALS.

The compositions described herein can be administered concomitantly withother non-omega-3 fatty acid medications prescribed for treatingneurodegenerative diseases. Non-limiting examples of medications caninclude medications generally approved by a health regulatory body andprescribed by physicians for a particular neurodegenerative disease. Incertain embodiments, the amount of the non-omega-3 fatty acid activeagent(s) administered in conjunction with the compositions describedherein can be reduced to avoid or minimize any side effects resultingfrom the non-omega-3 fatty acid active agents when such agents areadministered alone at higher concentrations.

Methods for Treating Primary Sclerosing Cholangitis

Methods are provided of treating PSC and or NSC in a subject in need oftreatment, which method comprises administering to said subject,typically a human adult, infant, child or adolescent, a therapeuticallyeffective amount of a composition of any one of the embodiments providedherein, or pre-formed micelles according to the respective embodimentsdescribed herein.

PSC, which is a chronic cholestatic liver disease, can result in fatmalabsorption syndromes. Thus, in certain embodiments, the compositionsdescribed herein can be administered to a human in need of suchadministration for the treatment of fat malabsorption resulting fromPSC.

In certain embodiments, the compositions described herein can beadministered in conjunction with non-omega-3 fatty acid active agents.Examples of such active agents can, upon approval by a respectiveregulatory agency, include 6-alpha-ethylchenodeoxycholic and/or(4R,5R)-1-[[4-[[4-[3,3-Dibutyl-7-(dimethylamino)-2,3,4,5-tetrahydro-4-hydroxy-1,1-dioxido-1-benzothiepin-5-yl]phenoxy]methyl]phenyl]methyl]-4-aza-1-azoniabicyclo[2.2.2]octanechloride.

In certain embodiments, the amount of the non-omega-3 fatty acid activeagents administered in conjunction with the compositions describedherein can be reduced to avoid or minimize any side effects resultingfrom the non-omega-3 fatty acid active agents when such agents areadministered alone at higher concentrations.

Methods for Treating Sickle Cell Disease

Methods are provided of treating sickle cell disease in a subject inneed of treatment, which method comprises administering to said subjecta therapeutically effective amount of a composition of any one of theembodiments provided herein, or pre-formed micelles according to therespective embodiments described herein.

The compositions described herein can be administered concomitantly withother non-omega-3 fatty acid active agents and non-omega-3 nutritionalsupplements prescribed for treating sickle cell disease. Non-limitingexamples of non-omega-3 active agents can include pain medications,antibiotics, hydroxyurea, anti-inflammatory medications, and aspirin,particularly a low-dose (70-81 mg) aspirin dosage form; whilenon-omega-3 nutritional supplements may include one or more of folicacid, magnesium, manganese, zinc, calcium, vitamin A, vitamin E, vitaminD, and vitamin K2. In certain embodiments, the amount of the non-omega-3fatty acid active agent(s) administered in conjunction with thecompositions described herein can be reduced to avoid or minimize anyside effects resulting from the non-omega-3 fatty acid active agentswhen such agents are administered alone at higher concentrations.

While not wishing to be bound to any particular theories, it is believedthat compositions comprising omega-3 DPA fatty acid (either in thetriglyceride, ester or free fatty acid ester form) and at least onesurface active agent in an amount and a combination effective to causesaid compositions to self-micellize when in contact with an aqueousmedium can be used to treat sickle cell disease symptoms, such asvaso-occlusive events, sequestration crisis, avascular necrosis, stroke,and sickle cell anemia, among other known maladies, by decreasing theaggregation of platelets, red blood cells, and/or white blood cells.

Nutritional Factors Associated with SCD Pathophysiological and ClinicalOutcomes

(A) Long Chain Polyunsaturated Fatty Acids (LCPUFA)

Until recently, cell membrane lipids (fatty acids) were regarded asbiologically inert compounds that provide a permeability barrier betweeninterior and exterior compartments within and between cells. It is nowrecognized that lipids, independently and in concert with proteins, arecentral to the regulation of cell and sub-cellular functions includingsignaling and gene expression. A third of all proteins are in membranes.These proteins, which are responsible for signaling, transport andoxidative protection and other vital cellular processes, are dependenton membrane lipids for their optimal function. Experimental and humanstudies demonstrate that abnormal composition, perturbation or deviationfrom normality of lipids leads to cell dysfunction.

Studies have shown that variation in dietary composition of saturatedand monosaturated fatty acids cause very little change in percentsaturation of membrane lipids. In contrast, membrane lipids are veryresponsive to variation to dietary composition of LCPUFA. The greatestsensitivity to changes in dietary fatty acids was for n-3 PUFA andn-6/n-3 fatty acids ratio. A 300% increase in red blood DHA and EPAcomposition has been observed in subjects supplemented with 1296 mg EPAand 864 mg DHA. Similarly, a 10 fold increase in EPA concentration ofmononuclear cell membrane phospholipids has been observed four weeksafter supplementation. Studies using multiple doses of fish oil showthat the incorporation of these fatty acids in immune cells occurs in amanner that is highly correlated with the amount of the fatty acidconsumed. Typically the increase in content of n-3 PUFAs occurs at theexpense of n-6 PUFAs, especially arachidonic acid.

Unlike n-6 fatty acids, n-3 fatty acids do not occur in large amounts inplants food and western human diet. Moreover, the alpha-linolenic acid(ALA) conversion process to EPA and DHA in humans is not efficient asonly 5-10% are converted to EPA, and a mere 2-5% to DHA. N-3 fatty acidsare abundantly present in fish and shellfish. In fact, fish-oilsupplements typically contain 30-50% of n-3 FAs. The current adequateintake (AI) for ALA issued by the Institute of Medicine of the NationalAcademies, USA is 1.6 g/day for men 19-47 years and 1.1 g/day for women19-47 years. The acceptable macronutrient distribution range (AMDR) forALA is 0.6-1.2% of energy. The lower boundary of the range meets the AIfor ALA. Approximately 10% of the AMDR for ALA can be consumed as EPAand/or DHA. The dietary guidelines (2005) also note that consumption ofapproximately two servings of fish per week (approximately 224 g total)may reduce the risk of mortality from coronary heart disease. However,since the physiological potency of EPA and DHA is greater than ALA andthere has been a substantive increase in the evidence base about thehealth benefits of long chain omega-3 fatty acids.

Perturbation of cell membrane fatty acids composition has been observedon red blood cells, platelets and mononuclear cells of patients withSCD. The abnormality is characterized by high omega-6, low omega-3 andan imbalance between the two fatty acid families. The n-3 and n-6 LCPUFAare vital structural and functional components of cell and sub-cellularcomponents. Studies have shown that the balance between these two fattyacid families influences blood cell adhesion, aggregation, bloodcoagulation, cell deformability and inflammatory response. Hence, it hasbeen postulated that an imbalance in membrane n-6/n-3 LCPUFA is theantecedent of the loss of membrane asymmetry, blood cell adhesion andaggregation and vaso-occlusion in SCD. Interestingly, experimental andclinical studies have shown that supplementing SCD patients with omega-3fatty acids corrects cell membranes abnormalities and confer protectionagainst vaso-occlusive pain episodes, severe anaemia and oxidativestress, and improves red blood cells flexibility in mice, modulates theeicosanoids and resolvin production, expression of adhesive molecules,and the overall inflammatory state. The retroconversion of DHA tosatisfy the omega-3 deficiencies at this cellular level results in anormalization of the cell membrane and alleviation of symptoms of thedisease state. The displacement of ARA during this process decreasessystemic inflammation.

(B) Antioxidants

Patients with sickle cell disease are under oxidative overload caused byaccelerated auto-oxidation and iron de-compartmentalization. Among thefactors thought to participate in systemic high oxidative stress (OS)associated with SCD is the increased intravascular haemolysis andexcessive levels of cell-free haemoglobin with its catalytic action onoxidative reaction. In addition, a chronic pro-inflammatory state is acharacteristic of SCD patients even in steady state, and reactive oxygenspecies (ROS) production is a main feature of and plays an importantrole in inflammation. It has also been demonstrated thathypoxia-reoxygenation cycles in SCD are associated with increased OS andsevere reperfusion injury. On the other hand, it has been found that theanti-oxidant capacity in RBCs of SCD patients is highly impaired. Thus,the chronic OS state associated with SCD is an outcome of imbalancebetween enhanced generation of reactive oxygen species (ROS) anddisrupted antioxidant system. It has been postulated that the impairedantioxidant capacities could be due to an accelerated utilization ofantioxidant nutrients. There is evidence to indicate that patients withSCD have a reduced levels of vitamin A, C, E and beta carotene and zinc.

(C) Protein

Sickle cell disease is associated with hyper-metabolism and a consequentshortage of substrates for normal growth and healthy immune response.Insufficient nutrition in general is associated with poor immunefunction and is consequently regarded as the most common cause ofimmunodeficiency worldwide. The protein:energy ratio is a majordeterminant of dietary adequacy.

Experimental studies have demonstrated that high protein intake resultsin lower circulating inflammatory markers, fewer infarcts in spleen,liver, and kidney, and lower histopathologic scores for chronic tissueinjury in liver and spleen. In addition, high-protein fed sickle micehad less vascular leakage in the heart, lungs, and brain and a betteracute immune response and survival rate than matched normal-proteinsickle mice. These experimental studies suggest that that the differencein nutritional intake might be a major contributory factor that explainspartially the wide spectrum of SCD clinical manifestation and severity.However, to the best to our knowledge, no study has investigated thelong-term effect of low or high protein and energy intake on SCD acuteand chronic complications.

Based on the previous studies on nutritional modifiers of sickle celldisease and the FDA approved Daily Reference Values (DRVs) and ReferenceDaily Intakes (RDIs) for adults and children four or more years of age,the following dietary formula to prevent acute and chronic complicationsassociated with SCD:

Protein: 50 g Vitamin A: 5,000 International Units (IU) Vitamin C: 60 mgVitamin E: 30 IU

Beta carotene:

Folate: 400 μg Zinc: 15 mg Manganese: 2 mg Copper: 2 mg Selenium: 70 μg

Omega-3 fatty acids (DHA AND EPA): 500 mg

Kits

Packaged pharmaceutical kits comprising the compositions describedherein are contemplated. The kits comprise the compositions describedherein as unit dosage forms in a container made of an inert material,such as for example, a glass vial, which can be clear or colored. Theglass vial can be a crimp or snap top glass vial. Included in the kitsare instructions for using the dosage form to treat a subject having adisease or disorder responsive to treatment by administration of thedosage forms comprising the compositions described herein.

The packaged pharmaceutical kits provide prescribing information, overthe counter medical use information, and/or nutritional information forthe dosage form including, for example and without limitation, to asubject or health care provider, or as a label in a packagedpharmaceutical kit. Information included in the kit may include, forexample and without limitation, efficacy, dosage and administration,contraindication and adverse reaction information pertaining to theomega-3 fatty acid compositions described herein. The dosage andadministration information, for example, can include dosing frequencyrecommendations as well as administration of the compositions with orwithout food.

In certain embodiments the dosage forms comprising the compositionsprovided herein are in the form of a liquid filled or gel filledcapsules for oral administration, which are provided either as blisterpackages or in bottles together with over the counter or prescriptionmedical use information and/or nutritional information.

The packaged pharmaceutical kits can comprise one or more of the omega-3fatty acids comprising the compositions described herein as the onlyactive ingredient. In other embodiments, one or more of the compositionsdescribed herein can be packaged in combination with one or more activeagents other than a non-Omega 3 fatty acid, such as for example andwithout limitation, one or more other lipid lowering or cholesterollowering agents selected from the group consisting of cholesterolabsorption inhibitors, bile acid sequestrants/resins, statins, niacinand derivatives, MTP inhibitors, fibrates and CETP inhibitors,6-alpha-ethylchenodeoxycholic and/or(4R,5R)-1-[[4-[[4-[3,3-Dibutyl-7-(dimethylamino)-2,3,4,5-tetrahydro-4-hydroxy-1,1-dioxido-1-benzothiepin-5-yl]phenoxy]methyl]phenyl]methyl]-4-aza-1-azoniabicyclo[2.2.2]octanechloride, insulin sensitizers, hypolipidemics, anti-inflammatorymedications, and thiazolidinediones, pain medications, antibiotics,folic acid, hydroxyurea, and anti-inflammatory medications.

In certain embodiments such active agents can include medicationsgenerally approved by a health regulatory body and prescribed byphysicians for a particular neurodegenerative disease.

In other embodiments, one or more of the compositions described hereincan be packaged in combination with one or more nutritional supplements.Non-limiting examples of supplements include various minerals, such ascalcium, magnesium, iron, and vitamins, particularly fat-solublevitamins, as well protein supplements well known in the art.

Dosage Forms

Any of the omega fatty acid containing compositions provided herein canbe provided as a pharmaceutical composition to be administered orally orparenterally, as a nutraceutical formulation, or a dietary supplement.

In certain embodiments of the invention, compositions comprise at leastone surface active agent, at least one fat-soluble vitamin, such asvitamin A, vitamin D, vitamin K, vitamin K2, among others, and anomega-3 fatty acid (in triglyceride, ester, or free fatty acid form). Incertain embodiments of the invention, at least one surface active agentis combined with at least one fat-soluble vitamin and an omega-3 fattyacid (in triglyceride, ester, or free fatty acid form) in relativeconcentrations sufficient to form micelles when added to an aqueousmedium. In certain other embodiments of the invention, at least onesurface active agent is combined with at least one fat-soluble vitamin,an omega-3 fatty acid (in triglyceride, ester, or free fatty acid form),and one or more minerals, such as magnesium, manganese, zinc, copper,selenium, and combinations thereof.

In certain embodiments of the invention, at least one surface activeagent is combined with at least one fat-soluble vitamin, an omega-3fatty acid (in triglyceride, ester, or free fatty acid form), and one ormore minerals, such as magnesium, manganese, zinc, copper, selenium, andcombinations thereof, such that when added to an aqueous medium, thecomposition forms micelles and wherein said micelles range in size from1 um to 10 um in diameter.

The pharmaceutical compositions described herein may further include oneor more pharmaceutically acceptable excipients. Pharmaceuticallyacceptable excipients include, but are not limited to, carriers,preservatives, and/or coloring agents. General considerations in thecomposition and/or manufacture of pharmaceutical compositions may befound, for example, in Remington The Science and Practice of Pharmacy21st ed., Lippincott Williams & Wilkins, 2005.

In certain embodiments, the compositions described herein can beformulated as a liquid for oral administration. Liquid compositionsinclude solutions, suspensions and emulsions. Examples of liquidpharmaceutical preparations include propylene glycol solutions andsolutions containing sweeteners for oral solutions, suspensions andemulsions. When the liquid composition comes into contact with anaqueous medium, such as for example an aqueous medium having an acidicenvironment, the composition forms micelles.

In certain embodiments, the dosage form comprises micelles which may bepre-formed prior to administration to a subject in need of suchadministration, and wherein said micelles range in size from 1 um to 10um in diameter. Such pre-formed micelles are stable at room temperature.

In other embodiments, the compositions described herein can beformulated as a fill material for capsules, including for example, asoft gelatin capsule. Likewise, when the contents of the soft gelatincapsule comes into contact with an aqueous medium, the composition formsmicelles upon disintegration of the capsule.

A capsule may be prepared, e.g., by placing the compositions describedabove inside a capsule shell. A capsule is a dosage form administered ina special container or enclosure containing an active agent. In someembodiments the compositions described herein can be filled into softcapsules. A capsule shell may be made of methylcellulose,hydroxypropylmethyl cellulose, polyvinyl alcohols, or denatured gelatinsor starch or other material. Hard shell capsules are typically made ofblends of relatively high gel strength bone and pork skin gelatins. Insome embodiments the unit dosage form is a gel capsule. In someembodiments the capsule shell is a glycerin capsule shell, for exampleproduct no. GSU0051 manufactured by SwissCaps and which meets USP 25requirements (SwissCaps, USA 14193 SW 119th Ave., Miami/Fla., U.S.33186). In other embodiments the capsule is a bovine gelatin shell, forexample SwissCaps product no. GSU0708. Other suitable capsule shellmaterials include polyethylene, polypropylene, poly(methylmethacrylate),polyvinylchloride, polystyrene, polyurethanes, polytetrafluoroethylene,nylons, polyformaldehydes, polyesters, cellulose acetate, andnitrocellulose. The capsule shell itself may contain small amounts ofdyes, opaquing agents, plasticizers, and preservatives. Conventionalmethods for preparing other solid dosage forms, for example, capsules,suppositories, and the like are also well known. Gelatin capsule shellsmay be made also be made of tapioca, grass, vegetable derived or fishderived gelatin. For example K-CAPS (Capsuline, Inc. Pompano Beach,Fla.) is a certified Kosher soft capsule shell of vegetable origin.Other vegetarian derived gelatin capsules may, be made of vegetablederived hydroxypropylmethyl cellulose (HPMC). Capsules shells may alsocontain Modified Maize Starch, Glycerol, and Carrageenan as a gellingagent.

In other embodiments the capsule has a shell comprising the material ofthe rate-limiting membrane, including coating materials, and filled withthe compositions described herein. Capsule shells may be made of aporous or a pH-sensitive polymer made by a thermal forming process. Incertain embodiments the capsule shell in the form of an asymmetricmembrane; i.e., a membrane that has a thin skin on one surface and mostof whose thickness is constituted of a highly permeable porous material.

Yet another useful capsule, a “swelling plug device”, can be used. Thecompositions described herein can be incorporated into a non-dissolvingcapsule-half of the device which is sealed at one end by a hydrogelplug. This hydrogel plug swells in an aqueous environment, and, afterswelling for a predetermined time, exits the capsule thus opening a portthrough which the active agent can leave the capsule and be delivered tothe aqueous environment. Preferred hydrogel-plugged capsules are thosewhich exhibit substantially no release of active agent from the dosageform until the dosage form has exited the stomach and has resided in thesmall intestine for about 15 minutes or more, preferably about 30minutes or more, thus assuring that minimal omega-3 fatty acid (eitherin the triglyceride, ester, or free fatty acid form) is released in thestomach or the small intestine. Hydrogel-plugged capsules of this typehave been described in patent application WO90/19168.

The dosage forms may contain a plasticizer, such as for example, in acapsule shell. Suitable plasticizers include, e.g., polyethylene glycolssuch as PEG 300, PEG 400, PEG 600, PEG 1450, PEG 3350, and PEG 800,stearic acid, propylene glycol, oleic acid, triethyl cellulose,triacetin, glycerin, sorbitol, sorbitan or combinations thereof.

In additional embodiments, the compositions can be formulated as aliquid for parenteral administration, particularly for intravenousadministration, added to infant formula, or injected into an intravenousbag comprising a saline or other pharmaceutically acceptable solution ora nutritional supplement.

The compositions described herein can be formulated as one or moredosage units. In some embodiments, it can be advantageous to formulateoral compositions in dosage unit form for ease of administration anduniformity of dosage. Dosage unit forms described in some embodimentscan refer to physically discrete units suited as unitary dosages for thesubject to be treated; each unit containing a predetermined quantity ofactive composition calculated to produce the desired therapeutic effectin association with the suitable pharmaceutical carrier.

In certain embodiments, the dosage form may optionally contain aflavorant such as orange oil, substantially pure d-limonene, and anantioxidant such as tocopherol, tocotrienol, ascorbyl palmitate or acombination of antioxidants.

Functional Foods

Certain embodiments of the present invention provide for functionalfoods comprising compositions which contain at least one surface activeagent and at least one omega-3 fatty acid (in triglyceride, ester, orfree fatty acid form) in relative concentrations sufficient to formmicelles when added to an aqueous medium, and wherein said micellesrange in size from 1 um to 10 um in diameter. These functional foods mayfurther contain one or more of (1) at least one fat-soluble vitamin,such as vitamin A, vitamin E, vitamin D, vitamin K, vitamin K2, amongothers, (2) one or more minerals, such as magnesium, manganese, zinc,copper, selenium, and combinations thereof, and (3) an omega-6, forexample arachidonic acid (ARA), linoleic acid (LA), gamma-linolenic acid(GLA) or the omega-6 isomer of DPA.

In certain embodiments, the compositions described herein comprisemicelles pre-formed prior to administration to a subject in need of suchadministration. Such pre-formed micelles are stable at room temperature.Accordingly, either such pre-formed micelles or said pre-micellizedcompositions in combination with a foodstuff, are provided, and can thenbe consumed as part of a healthy diet for enriching a subject's omega-3fatty acid levels or as a dietary treatment in addition to theoral/parenteral administration of the compositions described herein asprescribed by a health professional.

In certain embodiments, the functional food is in the form of edible ordrinkable compositions, e.g., foodstuffs such as chewable or ediblebars, confectionary products (e.g., chocolate bars), cookies, juicedrinks, baked or simulated baked goods (e.g., brownies), biscuits,lozenges or chewing gum. Examples of chewable or edible bars includechocolate bars or energy bars. Such functional foods can be particularlyuseful to people participating in sports or other forms of exercise.

In certain embodiments, the functional foods may also be in the form of,for example, butter, margarine, bread, cake, milk shakes, ice cream,yogurt and other fermented milk product.

In certain embodiments, the functional food can also be in the form of aliquid to be sprayed on meats, salads or other foods.

Other forms of the functional foods can be baby food, breakfast cereals,such as for example, grain flakes, muesli, bran, oatmeal.

When the functional food product is in a drinkable form, thecompositions described herein can be added directly to the drink, suchas for example plain milk, flavored milk, fermented milk products orjuices, or infant formula. The compositions will form micellescomprising the omega-3 fatty acids in the drinkable product.

When the functional food is in the form of a solid edible product, thecompositions described herein can be first added directly to afunctional food or to an aqueous medium, wherein the composition willform micelles as described herein. The aqueous medium comprising themicelles can subsequently be either sprayed onto the solid edibleproduct or mixed into the ingredients when manufacturing the edibleproduct.

In certain embodiments particularly for the treatment of sickle celldisease, at least one surface active agent and one or more omega-3 fattyacids (in triglyceride, ester, or free fatty acid form, e.g. DHA aloneor DHA and EPA: 500 mg), are further combined with Protein: 50 g,Vitamin A: 5,000 International Units (IU); Vitamin C: 60 mg, Vitamin E:30 IU, Beta carotene, Folate: 400 μg, Zinc: 15 mg, Manganese: 2 mg,Copper: 2 mg, and Selenium: 70 μg.

In certain embodiments of the invention, at least one surface activeagent is combined with at least one fat-soluble vitamin and an omega-3fatty acid (in triglyceride, ester, or free fatty acid form) in relativeconcentrations sufficient to form micelles when added to an aqueousmedium. In certain other embodiments of the invention, at least onesurface active agent is combined with at least one fat-soluble vitamin,an omega-3 fatty acid (in triglyceride, ester, or free fatty acid form),and one or more minerals, such as magnesium, manganese, zinc, copper,selenium, and combinations thereof.

EXAMPLES Macular Degeneration Example 1

The amounts and percentages of the ingredients comprising one embodimentof the compositions described herein are shown in Table 1:

TABLE 1 COMPOSITION INGREDIENT Amount (mg) % (wt/wt) DHA Omega-3 fattyacid Ethyl 754.3 68.57 Ester*† Polysorbate 80 337.9 30.72 Poloxamer 237(Pluronic ® F87) 7.8 0.71 TOTAL 1100 100 *The omega-3 oil may contain~2% α-tocopherol as an antioxidant. †DHA comprises at least 90% of theDHA oil (678.9 mg). The majority of the remaining no more than 10% isEPA (~5%, or up to 37.7 mg)

Example 2

The amounts and percentages of the ingredients comprising one embodimentof the compositions described herein are shown in Table 2:

TABLE 2 COMPOSITION INGREDIENT Amount (mg) % (wt/wt) EPA/DHA Omega-3fatty acid 754.3 65.59 Ethyl Ester*† Gamma-tocotrienol 50.0 4.35Polysorbate 80 337.9 29.38 Poloxamer 237 (Pluronic ® F87) 7.8 0.68 TOTAL1150 100 *The Omega-3 oil may contain ~2% α-tocopherol as anantioxidant. †EPA comprises 383 mg and 170 mg DHA.

Malabsorption Syndrome Example 3

A study was conducted to evaluate, in an established porcine model ofSBS, the systemic absorption and intestinal adaptation capacity of anethyl ester form of DHA in combination with a surface active agentcomposition effective to spontaneously form micelles with said fattyacids upon contact with an aqueous media. In certain embodiments, theself-micellizing compositions may further contain at least oneadditional omega-3 fatty acid (either in the triglyceride, ester, orfree fatty acid form), for example EPA or other known omega-3 fattyacids as disclosed herein. In certain embodiments, the self-micellizingcompositions may further contain an anti-oxidant, such as tocopherol. Incertain embodiments, the self-micellizing compositions may furthercontain an omega-6 fatty acid (either in the triglyceride, ester, orfree fatty acid form), for example arachidonic acid (ARA), linoleic acid(LA), gamma-linolenic acid (GLA) or the omega-6 isomer of DPA, alsoknown as osbond acid. This composition allows for micelle formationindependent of the presence of bile salts as compared to a standardtriglyceride form of DHA without a surface active agent (Control).

Materials and Methods

Animals and Surgical Induction of Short Bowel Syndrome

Newborn, crossbred (Hampshire×Landrace×Duroc×Yorkshire) piglets (n=10),were weighed, placed in cages in a heated room (˜30° C.), and were fedwith a commonly available sow milk replacement formula. After anovernight fast, piglets underwent surgery for the placement of jugularand intragastric catheters for postoperative parenteral and enteralnutrition, respectively, and small bowel resection surgery. Following a4-cm midline incision an 80% mid-jejunoileal resection was performed.The total amount of bowel resection was approximated using the followingequation: total intestinal length=280 cm×body weight{circumflex over( )}0.60. A 20% remnant (˜60 cm total per kg BW) remained and wasrepresented equally between proximal jejunum and ileum, representing ˜30cm each remnant segment per kg bodyweight. ˜45 cm of jejunum distal tothe ligament of Treitz and ˜45 cm of ileum proximal to the ileocecaljunction were measured using sterile silk ribbon placed along theantimesentric border of the gently stretched small intestine. Intestinenot included in the measurement was removed via cauterization. Bowelcontinuity was restored using an end-to-end jejunoileal anastomosis.

Postoperative Nutrition

Parenteral nutrition formula which does not contain DHA was providedproportionately to body weight to all piglets within 3 h after theoperation. Continuous enteral feeding with a commonly available sow milkreplacer formula, also lacking DHA, was slowly introduced 24 h afterintestinal resection at 3 mL/kg*h via the intragastric catheter. Thevolume of enteral feedings was then advanced every 24 h by 1 ml/kg*huntil a final enteral volume of 5 mL/kg*h was achieved on postoperativeday 3 which represents approximately 50% of full enteral feeding volumeand nutrient intake.

Provision of Oral Treatment & Control Groups

After the surgical procedure, piglets were randomly allocated into twogroups (N=5 per group), control and SBS-treatment group. The controlgroup received a standard DHA ethyl ester preparation of substantiallypure DHA ethyl ester (>90% DHA ethyl ester). The SBS-treatment groupreceived an equivalent amount of DHA as the control group, based onanimal weight as described herein, of a preferred embodiment of thepresent invention comprising, about 683.9 mg/g of substantially pure DHAethyl esters (>90% DHA ethyl ester), 2.7 mg/g of alpha-tocopherol, 306.3g/mg of polysorbate 80, and 7.1 mg/g of poloxamer 237.

SBS-treatment: The control and SBS-treatment formulations, respectively,mixed in the enteral feeding at the respective dose and infused via theintragastric catheter at a dose of 1 g/kg*day for a period of 4 daysafter which time the animals were sacrificed.

Growth Data and Sample Collection

Growth

Piglets were weighed every other day. In addition to calculating achange in weight by subtracting the starting weight at the time ofrandomization from the final weight, percent weight change and growthvelocity (g/kg*d) were also calculated. Percent weight change wascalculated by: [((final weight-starting weight)/starting weight)×100].Growth velocity was calculated by: [(weight change/starting weight)/4days of treatment].

TABLE 3 Cohort characteristics and weight parameters (median ± IQR)Control (n = 5) Treated (n = 5) p Male, n (%) 4(80) 3(60) 0.51 Weight atstart, g 2490 ± 114  2474 ± 325  0.75 Weight at finish, g 2594 ± 170 3170 ± 800  0.35 Change in weight, g 132 ± 278 696 ± 475 0.08 Growthvelocity, g/kg/d 13.5 ± 30.1 69.9 ± 41.5 0.08 Weight change, %  5.4 ±12.1 28.0 ± 16.6 0.08

Longitudinal Plasma Fatty Acid Levels

Blood samples for systemic fatty acid profiles were obtained on the dayof surgery and then twice daily until sacrifice on fourth postoperativeday. Plasma fatty acids were isolated and measured.

Intestinal Samples and Morphometry Measurements

Samples of intestinal tissue resected at the time of surgery andremnants at sacrifice were collected from proximal jejunum (proximal toanastomosis) and distal ileum (distal to anastomosis) and morphologicalmeasurements of the crypts and villi were measured. Only intact villiand crypts were measured. All measurements were made by the same,single, blinded observer.

Statistical Analysis

The primary outcome measure was a change in plasma DHA level in theSBS-treated group versus control group. Given the small number ofpiglets per group, appropriate small group statistical methods were usedto evaluate differences in outcome variables. Variable summarystatistics are expressed as median±interquartile range (IQR). TheWilcoxon Rank-Sum test, which is appropriate to compare two non-normaldistributions as is common in small sample size, was used to compareoutcome measures between groups; while the Wilcoxon signed-rank test,which is appropriate for repeated measurements within the same animal,was used to compare paired, before and after surgery outcome measureswithin groups. Differences in growth measurements and fatty acid levelsover time between groups were evaluated using generalized linear mixedmodels (GLMM) to account for repeated measures and non-Gaussian,non-normal data.

Results

Piglet Characteristics and Weight Trends

Four of the 5 piglets were male in the control group while 3 of the 5were male in the SBS-treatment group (Table 1). The median startingweight was similar in both groups. At the end of the protocol, themedian difference in final weight from starting weight was 696±425 g inthe SBS-treatment group compared to 132±278 g in the control group. Inconcordance with the weight difference the percent weight change andgrowth velocity was greater in the SBS-treatment group compared to thecontrol group. However, due to the short duration of this model as wellas the sample size, these differences in growth parameters did not reachstatistical significance.

Small Bowel Resection and Final Organ Weights

There were no differences in the total length or weight of resectedbowel per kg weight of the piglet (Table 4). A median total of 138±34 cmper kg of small bowel was removed from piglets in the SBS-treatmentgroup compared to 119±7 cm per kg in the control group. Similarly, therewere no differences in the final length or weight of the jejunum andileum segments of the small bowel per kg weight. Finally, the liver andbrain weights per kg at sacrifice were comparable between the twogroups.

TABLE 4 Small intestine resection and final organ weights (median ± IQR)SBS- Control Treatment p Resected SI, g/kg 29.5 ± 2.2 28.7 ± 4.9 0.92Resected SI, cm/kg  119 ± 7.0  138 ± 3.4 0.17 Jejunum, g/kg  5.2 ± 1.4 5.1 ± 0.5 0.92 Jejunum, cm/kg 21.2 ± 6.7 16.5 ± 3.8 0.60 Ileum, g/kg 5.3 ± 1.2   7 ± 1.4 0.05 Ileum, cm/kg 17.5 ± 4.7 19.2 ± 4.4 0.33 Liver,g/kg 36.3 ± 2.5 37.0 ± 4.2 0.71 Brain, g/kg 14.0 ± 1.6 11.6 ± 2.3 0.09IQR: Interquartile range; SI: Small Intestine; SBS-treatment

Plasma Fatty Acid Levels

Initial plasma fatty acid levels in mol % (Time 0) of DHA,Eicosapentaenoic acid (EPA), ARA, and Linoleic acid (LA) obtained at thetime of surgery were similar between groups (2.0±0.04 vs 2.3±0.4;0.2±0.06 vs 0.3±0.1; 11.0±0.3 vs 11.8±0.5; 18.1±2.3 vs 16.6±1.8,respectively) (FIG. 1). Within 12 hours (after the first enteral dose ofDHA), the DHA and EPA levels were significantly greater in theSBS-treatment group versus control group (4.1±0.3 vs 2.5 vs 0.5,p=0.009; 0.7±0.3 vs 0.2±0.005, p=0.009, respectively). For DHA thesedifferences persisted until the final measurement; while the EPA plasmalevel differences persisted throughout the remaining duration of theprotocol. On the third postoperative day, plasma ARA levels began todiverge between the groups, with lower values in the SBS-treatment groupversus the control group (7.1±1.5 vs 8.7±0.8, p=0.03). The differencepersisted throughout the remaining duration of the protocol. There wereno differences throughout the study protocol in plasma LA levels.

Small Bowel Morphometry

At the time of surgery (Day 0), villus height measurements of theproximal jejunum and distal ileum were similar in both groups (FIG. 2).In the control group, the villus height of the proximal jejunum at theend of the experiment (Final) was not significantly different to thevalues obtained at the time of surgery; whereas in the distal ileumvillus height was reduced, but not significantly different (p=0.07). Incontrast, in the SBS-treatment group, the final villus height wasincreased from the starting value in the proximal jejunum (p=04). In thedistal ileum, the SBS-treatment group demonstrated maintenance ofoverall villus height and compared to the control group, a significantincrease in the final villus height (p=0.01).

Crypt depth was increased at the end of the experiment compared tobaseline values in the proximal jejunum within both control andSBS-treatment groups (p<0.05). There were no differences between groupsin the baseline or final crypt depth measurements. Similar to theproximal jejunum, the crypt depth of the distal ileum was increased frombaseline in the distal ileum within the control group (p<0.05). Incontrast, there were no changes in the baseline to final crypt depth inthe distal ileum for the piglets in the SBS-treatment group. The finalcrypt depths in the distal ileum were not significantly differentbetween the two groups.

Discussion

Piglets with surgically induced SBS who were given dietarysupplementation in the SBS-treatment group demonstrated improvedabsorption of DHA, greater ileal adaptation, and a trend towardsimproved growth compared to piglets given a standard preparation of DHA.Within 12 hours, the plasma levels of both DHA and EPA in theSBS-treatment group began to significantly diverge from the controlgroup. While not wishing to be bound to any particular theory, anincrease of EPA is believed to be a result of retro-conversion of DHA toEPA as neither DHA formulation contained an EPA supplement. The observedreduction in ARA with increasing DHA levels in the SBS-treatment groupis a known phenomenon. However, while not wishing to be bound to anyparticular theory, as DHA levels increase with DHA-only dietarysupplementation, ARA levels decline as a result of the competition forenzyme activity, the sn-2 position of phospholipids in membranes, aswell as the body's ability within the plasma compartment to maintain aconstant number of double bonds. This is not a generalized phenomenon asLinoleic Acid levels were unchanged throughout the experimental timeperiod.

An increase in villus height and crypt depth is a known observation inintestinal adaptation after significant bowel resection in both animalmodels and in humans. This was observed for both groups in the proximaljejunum and distal ileum reinforcing the validity of this model instudying intestinal changes in short bowel syndrome. In the proximaljejunum the SBS-treatment group demonstrated a greater attainment invillus height compared to baseline. In the distal ileum, theSBS-treatment group maintained villus height while the control grouplost villus height suggesting a greater intestinal adaptive response forthe piglets receiving the SBS-treatment preparation. The piglets in theSBS-treatment group appeared to have an accelerated adaptive responsecompared to the control group possibly due to the efficientbioavailability and intestinal absorption of DHA. This may be aclinically relevant observation during the critical, immediatepostoperative period. Although increased jejunal crypt depth was seen inboth groups after 4 days, the lack of change in the ileum selectively inthe SBS-treatment group suggests that increased bioavailability of DHAis exerting an intestinal trophic effect. While not wishing to be boundto any particular theory, it is possible that DHA is modulatingintestinal cell growth and differentiation through PPAR or other growthfactors.

The example described herein provides data necessary to support theclinical utility of a novel preparation of DHA that allows for bile acidindependent formation of micelles enhancing absorption and biologicaleffectiveness of DHA early in the postoperative period.

Example 4

The amounts and percentages of the ingredients comprising one embodimentof the compositions described herein are shown in Table 5:

TABLE 5 COMPOSITION INGREDIENT Amount (mg) % (wt/wt) DHA Omega-3 fattyacid Ethyl 685.7 68.57 Ester*† Polysorbate 80 307.2 30.72 Poloxamer 237(Pluronic ® F87) 7.1 0.71 TOTAL 1000 100 *The omega-3 oil may contain~2% α-tocopherol as an antioxidant. †DHA comprises at least 90% of theDHA oil (617.3 mg). The majority of the remaining no more than 10% isEPA (~5%, or up to 34.3 mg)

During administration, sterile saline solution is added to the vial andmixed to homogeneity to form a milky solution, which can then beadministered intravenously directly into a human in need of suchadministration, such as for example a neonate, added to infant formula,into an IV infusion bag, via a stomach feeding tube, or through aduodenal feeding tube.

Example 5

The amounts and percentages of the ingredients comprising one embodimentof the compositions described herein are shown in Table 6:

TABLE 6 COMPOSITION INGREDIENT Amount (mg) % (wt/wt) DHA Omega-3 fattyacid Ethyl 651.4 65.14 Ester*† Arachidonic Omega-6 Ethyl 34.3 3.43Ester*° Polysorbate 80 307.2 30.72 Poloxamer 237 (Pluronic ® F87) 7.10.71 TOTAL 1000 100 *The omega-3 oils may contain ~2% α-tocopherol as anantioxidant. †DHA comprises at least 90% of the DHA oil (586.3 mg). Themajority of the remaining no more than 10% is EPA (~5%, or up to 32.6mg) °The ARA Ethyl Ester is at 80% pure.

NAFLD/NASH Examples Example 6

The amounts and percentages of the ingredients comprising one embodimentof the compositions described herein are shown in Table 7:

TABLE 7 COMPOSITION INGREDIENT Amount (mg) % (wt/wt) DHA Omega-3 fattyacid Ethyl 754.3 68.57 Ester*† Polysorbate 80 337.9 30.72 Poloxamer 237(Pluronic ® F87) 7.8 0.71 TOTAL 1100 100 *The omega-3 oil may contain~2% α-tocopherol as an antioxidant. †DHA comprises at least 90% of theDHA oil (678.9 mg). The majority of the remaining no more than 10% isEPA (~5%, or up to 37.7 mg)

Example 7

The amounts and percentages of the ingredients comprising one embodimentof the compositions described herein are shown in Table 8:

TABLE 8 COMPOSITION INGREDIENT Amount (mg) % (wt/wt) EPA/DHA Omega-3fatty acid Ethyl 754.3 65.59 Ester*† Gamma-tocotrienol 50.0 4.35Polysorbate 80 337.9 29.38 Poloxamer 237 (Pluronic ® F87) 7.8 0.68 TOTAL1150 100 *The omega-3 oil may contain ~2% α-tocopherol as anantioxidant. †EPA comprises 383 mg and 170 mg DHA.

Example 8

An embodiment of the present invention, referred to as SC410 herein, isa composition consisting of about: 68.39% docosahexaenoic acid ethylesters (DHA), 0.27% alpha-tocopherol, 30.63% polysorbate 80, and 0.71%poloxamer 237. As disclosed herein, SC410 is preferably administeredenterally to a mammal, such as a human, and delivered within a softgelatin capsule although the routes of administration and deliveryinclude other known routes. A nine-week preclinical study of certainembodiments of the present invention was conducted on 24 male mice usinga mouse model (“NASH mice”) which progresses from NAFLD to NASH between5 to 6 weeks of age. Within the disease progression of this animalmodel, the animals reach NASH around week 6. Eight NASH mice were orallyadministered an olive oil vehicle in a volume of 2.5 mL/kg once dailyfrom 5 to 9 weeks of age (Control Group). A second group, the “SecondTreatment Group,” also interchangeably referred to herein as NAFLDGroup, composition 1-5 group, or composition 1-5 treatment, consisted ofeight NASH mice which were orally administered said olive oil vehiclesupplemented with SC410 once daily from 5 to 9 weeks of age. A thirdgroup, the “Third Treatment Group,” also interchangeably referred toherein as NASH Group, composition 1-6 group or composition 1-6treatment, consisted of eight NASH mice, which were orally administeredsaid olive oil vehicle supplemented with SC410 once daily from 6 to 9weeks of age. Non-fasting blood glucose was measured in whole blood andALT levels were measured from plasma. Liver total lipid-extracts wereobtained from right lobes by Folch's method.

FIG. 3 shows the mean body weight on the day of sacrifice of the micebetween each of the Control, Second Treatment Group and Third TreatmentGroup.

FIG. 4 shows α-SMA mRNA gene expression in mice in each of the control,NAFLD and NASH groups treated with either Vehicle or Composition 1 atthe end of the treatment period.

FIG. 5 shows plasma alanine transaminase (ALT) levels in mice betweeneach of the control, NAFLD and NASH group treated with either Vehicle orComposition at the end of the treatment period.

FIG. 6 shows the mean liver weight on the day of sacrifice of the micebetween each of the treatment groups and the control.

FIG. 7 shows the mean liver to body weight ratio on the day of sacrificeof the mice between each of the treatment groups and the control.

FIG. 8 shows whole blood glucose in mice between each of the control,NAFLD and NASH group treated with either Vehicle or Composition at theend of the treatment period.

FIG. 9 shows liver triglyceride levels in mice between each of thecontrol, NAFLD and NASH.

FIGS. 10 A and 10B show the mean individual EPA (A) and DHA (B) totallipid concentration-time profiles (baseline-adjusted change) after asingle dose of SC401 during fed and fasting conditions.

FIGS. 11A and 11B show the mean individual EPA (A) and DHA (B) freefatty acid concentration-time profiles (baseline-adjusted change) aftera single dose of SC401 during fed and fasting conditions.

FIG. 12 shows mean EPA and DHA total lipid plasma concentration profiles(μg/ml) (baseline-adjusted) after administration of a single dose (doseadjusted) of SC401 and Lovaza® in fasted conditions.

Note—N.S. In the Figures Stands for Non-Significant.

Body Weight

At the end of the experiment, it was determined that body weightgradually increased during the treatment period in all groups as wouldbe expected in growing mice. There was no significant difference in meanbody weight between the Control Group and the First Treatment Groupduring the treatment period although mean body weight on the day ofsacrifice of the First Treatment Group tended to be decreased comparedwith the Control Group. However, the mean body weight of the SecondTreatment Group was significantly lower than that of the Control Groupat day 28.

α-SMA

α-SMA, a marker of myofibroblast associated with the development ofliver fibrosis, mRNA expression levels tended non-significantly todecrease in both treatment groups as compared with the Control Group.

ALT Measurements

Although changes in ALT were not statistically significant between thegroups, some animals within the treatment groups exhibited a markedincrease in plasma ALT levels as compared to the control group. Sinceomega-3 administration has been shown in previous studies to elevateALT, this result would be expected and may be an indicator of theefficacy of delivery of the omega-3 to the liver.

Liver Weight

Additionally, there was no significant difference in mean liver weightbetween the Control Group and the First Treatment Group, whereas themean liver weight of the Second Treatment Group was lower than that ofthe Control group. When liver and body weight are compared collectivelyas a ratio, the Second Treatment Group non-significantly trends lowerthan the Control or First Treatment Group.

Blood Glucose

There was no significant difference in whole blood glucose between theControl Group and the First Treatment Group whereas whole blood glucoselevel tended to decrease in the Second Treatment Group compared with theControl Group.

Liver Triglycerides

Liver triglyceride contents tended to decrease in the First TreatmentGroup and significantly decreased in the Second Treatment Group comparedwith the Control Group.

CONCLUSIONS

The study showed that animals for which treatment was started later, theSecond Treatment Group, had significantly reduced the liver triglyceridecontents compared with the control group. Although the livertriglycerides tended to decrease in both treatment groups, the FirstTreatment Group showed a non-significant change (at 0.05) and the SecondTreatment Group showed a statistically significant decrease. This SecondTreatment Group also showed a decreasing tendency of whole blood glucoselevels compared with the Control Group. The significant decrease in theliver triglycerides in the more progressed group demonstrate thatcertain embodiments of the present invention can improve lipid andglucose metabolism in the liver. These results also demonstrate thatomega-3 fatty acids can improve lipid and glucose metabolism in theliver.

The effect of the treatment was not merely through an induction ofweight loss by the animals. In fact, the study found that body weightgradually increased during the treatment period in all groups and thatthere was no difference in mean body weight between groups. Although theend liver weight of the subjects in the treatment groups was lower thanthat of the subjects in the Control Group, the difference was notstatistically significant. In addition, both treatment groups trended toreduce alpha-SMA (a marker of myofibroblast associated with thedevelopment of liver fibrosis) mRNA expression in the liver, a keyindicator of the treatment's demonstrated ability to delay theprogression to NASH. There were no significant differences in fibrosisarea between the Control Group and either group, data not shown.

Without being held to any one theory, we believe the study demonstratesthe ability of at least one composition disclosed herein to deliverpolyunsaturated EPA and DHA fatty acids to mammals with NAFLD forincorporation into the cell membrane of erythrocytes, wherein saiddelivery can reduce the triglyceride level in the liver and thereby slowthe disease's progression to NASH. The study may also indicate thatadministration of compositions disclosed herein might provide a largermedicinal response when provided later in the disease progression fromNAFLD to NASH.

Neurodegenerative Diseases Prophetic Non-Limiting Working ExamplesExample 9

The amounts and percentages of the ingredients comprising one embodimentof the compositions described herein are shown in Table 9:

TABLE 9 COMPOSITION INGREDIENT Amount (mg) % (wt/wt) DHA Omega-3 fattyacid Ethyl 754.3 68.57 Ester*† Polysorbate 80 337.9 30.72 Poloxamer 237(Pluronic ® F87) 7.8 0.71 TOTAL 1100 100 *The omega-3 oil may contain~2% α-tocopherol as an antioxidant. †DHA comprises at least 90% of theDHA oil (678.9 mg). The majority of the remaining no more than 10% isEPA (~5%, or up to 37.7 mg)

Example 10

The amounts and percentages of the ingredients comprising one embodimentof the compositions described herein are shown in Table 10:

TABLE 10 COMPOSITION INGREDIENT Amount (mg) % (wt/wt) EPA/DHA Omega-3fatty acid Ethyl 754.3 65.59 Ester*† Gamma-tocotrienol 50.0 4.35Polysorbate 80 337.9 29.38 Poloxamer 237 (Pluronic ® F87) 7.8 0.68 TOTAL1150 100 *The omega-3 oil may contain ~2% α-tocopherol as anantioxidant. †EPA comprises 383 mg and 170 mg DHA.

Primary Sclerosing Cholangitis Prophetic Non-Limiting Working ExamplesExample 11

The amounts and percentages of the ingredients comprising one embodimentof the compositions described herein are shown in Table 11:

TABLE 11 COMPOSITION INGREDIENT Amount (mg) % (wt/wt) DHA Omega-3 fattyacid Ethyl 754.3 68.57 Ester*† Polysorbate 80 337.9 30.72 Poloxamer 237(Pluronic ® F87) 7.8 0.71 TOTAL 1100 100 *The omega-3 oil may contain~2% α-tocopherol as an antioxidant. †DHA comprises at least 90% of theDHA oil (678.9 mg). The majority of the remaining no more than 10% isEPA (~5%, or up to 37.7 mg)

Example 12

The amounts and percentages of the ingredients comprising one embodimentof the compositions described herein are shown in Table 12:

TABLE 12 COMPOSITION INGREDIENT Amount (mg) % (wt/wt) EPA/DHA Omega-3fatty acid Ethyl 754.3 65.59 Ester*† Gamma-tocotrienol 50.0 4.35Polysorbate 80 337.9 29.38 Poloxamer 237 (Pluronic ® F87) 7.8 0.68 TOTAL1150 100 *The omega-3 oil may contain ~2% α-tocopherol as anantioxidant. †EPA comprises 383 mg and 170 mg DHA.

Example 13

The study described below will assess the effect of compositionsdescribed herein in a murine model of Primary Sclerosing Cholangitis(PSC). It is believed that treatment with the compositions describedherein will be more effective in preventing and/or reversing bile ductinjury in a mouse model of PSC.

The study will utilize a mouse model of PSC induced by DSS & DDC. Thewell established mouse model of PSC where exon 10 CFTR^(+/−) knockoutmice (heterozygotes) will be exposed to oral Dextran Sodium Sulfate(DSS) followed by up to 28 days of the xenobiotic DDC dissolved inpeptamen (Martin C R, Zaman M M, Ketwaroo G A, Bhutta A Q, Coronel E,Popov Y, Schuppan D, Freedman S D. CFTR dysfunction predisposes tofibrotic liver disease in a murine model. Am J Physiol GastrointestLiver Physiol 303: G474-G481, 2012).

The Cftr^(+/−) mouse model of PSC consists of two phases: (1) inductionof colitis with DSS for 10 days or until overt blood per rectum; and,(2) induction of liver inflammation and fibrosis with DDC for 28 days.The following protocol will begin using Cftr^(+/−) mice at ˜age 35 days,or week 5.

Induction of Colitis with DSS for 10 Days

85 mg of DSS dissolved in 12 ml of Peptamen by continuous vortexing.

The mice will be fed using a liquid feeder (Dyets Inc., Bethlehem, Pa.).

The mice will be monitored daily for onset of colitis by overt bleedingper rectum.

Once overt bleeding is noted (typically day 8-10 of DSS), DSS will bediscontinued and the mice will be fed peptamen alone for one day.

Induction of Liver Inflammation and Fibrosis with Oral DDC for 28 Days

After overt colitis and one day of peptamen feeding alone, the mice willbe treated for 28 days with 1.5 mg DDC in 12 ml of Peptamen per day

Volume consumed will be recorded.

Treatment Groups

3 treatment groups will be used:

1) High DHA (40 mg/day)2) High DHA (40 mg/day) with high bioavailability3) No treatment

All treatments will begin 3 days prior to the administration of DSS andthus will examine prevention of injury.

Sample Collection and Outcomes Measures

a) Mice will be sacrificed by exposure to CO₂ for 2 min.b) Body weight will be measured at the start of the protocol, every3^(rd) day, then at the end of the protocol. Percent weight change willbe calculated as follows:i.=((Starting weight-End weight)/Starting weight)*100c) Liver and blood samples will be collected at sacrifice and stored asdescribed below in Table 13 below.d) Fatty acid levels will be determined in plasma and liver.

TABLE 13 Sample collection and processing Outcome Measure Sample Samplepreparation/storage Sample analysis Tissue Weight Liver Liver removedand weighed pre- BIDMC fixation Histology Liver Liver fixed in 10%buffered BIDMC for blind formalin for 2 hours followed by scoring twoPBS washes and stored in PBS at 4° C. H&E staining, Sirius red stainingFatty Acids Liver Fatty acid extraction BIDMC GC-MS Serum Blood will becollected by cardiac puncture using a 1 ml syringe equipped with a 25 Gneedle. Blood will be placed on ice for 15 minutes. The sample will becentrifuged at 5500Xg and serum collected. On average, 150-200 uL ofserum will be collected per mouse. The serum will be pre-aliquoted intoseparate vials for the following distribution. All aliquoted sampleswill be stored at −80° C. Fatty Acids Serum Fatty acid extraction BIDMCGC-MS The following will be collected and stored for possible subsequentanalyses depending on histology results. LOXL-2, α-SMA, or LiverImmunohistochemistry BIDMC other markers TBD Two separate pieces ofliver tissue will be provided for each animal. Tissue will be snap/flashfrozen in tissue biopsy cassettes using liquid nitrogen and will bestore at −80° C. Inflammatory/Fibrosis Liver Snap freeze, store at −80°C. RT PCR at BIDMC markers: procollagen-1, LOXL-2, TGF-β1, TGF-β2,TIMP-1, TIMP-2, MMP-2, MMP-9, TNF-α, α-SMA Liver Function Tests: SampleAmount Destination Alkaline Phosphatase, Serum 50 uL BIDMC ALT, AST,Bilirubin Luminex Cytokine Serum 70 uL BIDMC Panel

Tissue Histological Slide Scoring

a) Slides will prepared from the left lateral and bifurcated mediallobes of the liver; each liver will have one slide stained with H&E andone with Sirius Red respectively.b) Liver injury will be scored based on four criteria: (1) epithelialinjury, intraepithelial inflammation, and mononuclear cell infiltration;(2) bile duct proliferation; (3) bile duct angulation; and (4) fibrosis.Each criterion receives a score between 0 and 3. A scoring rubric isdefined in the box below. Scoring as previously described in our AJParticle, will be performed by two individuals blinded to the conditions

SCORING RUBRIC 0 Absent 1 Mild 2 Moderate 3 Severei. Epithelial injury, intraepithelial inflammation, and mononuclear cellinfiltration will be scored on slides at 20× magnification stained withH&E.ii. Bile duct proliferation will be scored at 5× magnification on slidesstained with H&E.iii. Bile duct angulation will be scored at 20× magnification on slidesstained with H&E.iv. Fibrosis will be scored on slides at 5× magnification stained withSirius Red. All tissues will be held and processed at the same time forSirius Red staining.

Fibrosis will be quantified on Sirius Red slides using ImageJ

Sickle Cell Disease Prophetic Non-Limiting Working Examples Example 14

The amounts and percentages of the ingredients comprising one embodimentof the compositions described herein are shown in Table 14:

TABLE 14 COMPOSITION INGREDIENT Amount (mg) % (wt/wt) DHA Omega-3 fattyacid Ethyl 754.3 68.57 Ester*† Polysorbate 80 337.9 30.72 Poloxamer 237(Pluronic ® F87) 7.8 0.71 TOTAL 1100 100 *The omega-3 oil may contain~2% α-tocopherol as an antioxidant. †DHA comprises at least 90% of theDHA oil (678.9 mg). The majority of the remaining no more than 10% isEPA (~5%, or up to 37.7 mg)

Example 15

The amounts and percentages of the ingredients comprising one embodimentof the compositions described herein are shown in Table 15:

TABLE 15 COMPOSITION INGREDIENT Amount (mg) % (wt/wt) EPA/DHA Omega-3fatty acid Ethyl 754.3 65.59 Ester*† Gamma-tocotrienol 50.0 4.35Polysorbate 80 337.9 29.38 Poloxamer 237 (Pluronic ® F87) 7.8 0.68 TOTAL1150 100 *The omega-3 oil may contain ~2% α-tocopherol as anantioxidant, †EPA comprises 383 mg and 170 mg DHA.

Example 16

A Randomized, double blind, placebo-controlled study will be conductedto investigate the therapeutic potential of the compositions describedherein for patients with homozygous sickle cell disease.

Objective:

-   -   1. To evaluate efficacy of the compositions described herein in        treatment of sickle cell anemia.    -   2. To study change in number of sickle cell vaso-occlusive        crises.    -   3. To monitor the safety of one or more of the compositions        described herein in the treatment of sickle cell anemia        -   Sample size: 60

Study medication (n=sample in each group):

-   -   1. Test (T): One or more of the compositions described herein        (n=30)    -   2. Reference (R): Standard of Care (n=30)

Duration of treatment: 6 months

Inclusion Criteria:

-   1. Patients aged 2-24 y with HbSS, who are undergoing regular    follow-up as an outpatient with Sickle Cell Disease.-   2. The patients who will be in a steady state, defined as no    evidence of fever, infection, or crisis for 4 wk before the start of    the study.-   3. Phenotypic characteristic was confirmed with the use of cellulose    acetate electrophoresis at pH 8.5.-   4. Can continue standard of care treatment.-   5. Already on fish oil compounds will require wash out of 15 days.

Exclusion Criteria:

-   1. Presence of other chronic diseases.-   2. Blood transfusion in the previous 4 mo.-   3. A history of overt stroke.-   4. Pregnancy.

Medication permitted: All of the patients receiving regular folatesupplementation, and those, 5 y receiving standard oral prophylacticpenicillin.

Investigation Parameters: Blood count, inflammatory markers (C-RP)

The primary end point will be rates of clinical vaso-occlusive crisisand secondary end points will include number of hospitalizations,haemolytic events, blood transfusion rate, school attendance, and bloodcount—will be analysed by intention-to-treat analysis and quality oflife assessment.

Statistics: N=30. The vaso-occlusive crises will be summarized on thebasis of the annualized crisis rate by dividing the total number ofcrises experienced by the number of follow-up months and multiplying by12.

Example 17

The amounts and percentages of the ingredients comprising one embodimentof the composition, also referred to as SC401, are shown in Table 17:

TABLE 17 INGREDIENT Amount (mg) % (wt/wt) COMPOSITION (FILL MASS)/dosageform Total Omega-3 fatty acid Ethyl Esters 754.3 68.57 EPA Ethyl Esters392.2 35.65 DHA Ethyl Esters 165.9 15.08 Polysorbate 80 337.9 30.72Poloxamer 237 (Pluronic ® F87) 7.8 0.71 GEL MASS/dosage form Gelatin 27040 Glycerin 135 20 Purified water 270 40

The pharmacokinetic (PK) profiles and bioavailability of EPA and DHAcomprising the SC401 formulation is graphically represented in FIGS.10A, 10B, 11A AND 11B.

FIG. 10 shows mean EPA and DHA total lipid plasma concentration profiles(μg/ml) (baseline-adjusted) after administration of a single dose (doseadjusted) of SC401 and Lovaza® in fasted conditions.

1. A method of treating or preventing sickle cell disease or diseasesymptoms, comprising administering an effective amount of a compositionto a subject in need thereof, the composition comprising: at least 90%pure docosahexaenoic acid (DHA) and at least one surface active agent,wherein said at least one surface active agent comprises a combinationof at least one polysorbate and at least one poloxamer, wherein said atleast one polysorbate comprises from about 30.0% wt/wt to about 32.0%)wt/wt of said composition, wherein said at least one poloxamer comprisesfrom about 0.60% wt/wt to about 0.80%) wt/wt of said composition,wherein said at least one polysorbate is polysorbate 80, and whereinsaid at least one poloxamer is poloxamer 237.